WO2009026248A2 - Hydrazide, amide, phthalimide and phthalhydrazide analogs as inhibitors of retroviral integrase - Google Patents

Abstract

The present invention provides catechol-containing hydrazides, amides, phthalimide and phthalhydrazide analogs. These compounds are inhibitors of retroviral integrase, an essential enzyme for the proliferation of retroviruses such as HIV-1. Also provided are pharmaceutical compositions comprising at least one of the catechol-containing hydrazides, amides, phthalimide or phthalhydrazide analogs and a method of using the hydrazide, amide, phthalimide and phthalhydrazide analogs to inhibit retroviral proliferation and as therapeutics for the treatment of AIDS.

Description

HYDRAZIDE, AMIDE, PHTHALIMIDE AND PHTHALHYDRAZIDE ANALOGS AS INHIBITORS OF RETROVIRAL INTEGRASE

BACKGROUND OF THE INVENTION

[0001] Retroviruses are RNA viruses that are capable of inserting their genomic sequence, following reverse transcription, into a host cell's genome. Numerous diseases in a variety of animal species are known to result from retroviral infection, including those by avian sarcoma virus (ASV), feline leukemia virus (FeLV), Moloney murine leukemia virus (MoMLV), and simian immunodeficiency virus (SIV). The most notable among human diseases that are attributable to retroviral infection include human T cell leukemia/lymphoma caused by the human T-cell lymphotropic virus (HTLV) and the acquired immunodeficiency syndrome (AIDS) caused by the human immunodeficiency virus (HIV).

[0002] Integration of reverse transcribed viral DNA into the host chromosome is an essential step in retroviral replication. This process is catalyzed by the viral enzyme integrase and establishes the integrated viral DNA (or the provirus) as a genetic component that persists throughout the life of the host cell. Because of the role of this enzyme, retroviral integrase has long been considered an important target in anti-retroviral drug design.

[0003] Inhibitors of HIV- 1 integrase (IN) have emerged as a promising new class of therapeutics for the treatment of AIDS (Palmisano, L. Exp. Rev. Anti-Infect. Ther. 2007, 5, 67-75). Although IN has long been regarded as a potentially attractive target for anti-HIV drug development, discovery of clinically-relevant inhibitors has been challenging (Hazuda, D. J. and Young, S. D. Adv. Antiviral Drug Des. 2004, 4, 63-77; Dayam, R. et al.. Med. Res. Rev. 2006, 26, 271-309; Pommier, Y. et al. Nature Rev. Drug. Discov. 2005, 4, 236-248). Many potent inhibitors against the IN-catalyzed 3 '-processing (3'-P) and strand transfer (ST) reactions were initially developed using in vitro IN assays (Craigie, R.et al. Nucleic Acid Res. 1991, 19, 2729-2734) that employ Mn²⁺ as a metal cofactor (Pommier, Y. and Neamati, N. Adv. Virus Res. 1999, 52, 427-458). However, under physiological conditions Mg²⁺ serves as the IN cofactor and frequently these inhibitors either failed to show good antiviral potencies in HIV-I infected cells or they inhibited by non-IN dependent mechanisms (Witvrouw, M.; et al.. Curr. Med. Chem. Anti-Infect. Agents 2005, 4, 153-165). The identification of metal chelating inhibitors bearing the general structure i (Figure 1) was a significant advance in the field of IN inhibitor design (Hazuda, D. J. and Young, S. D. Adv. Antiviral Drug Des. 2004, 4, 63-11; White, E. H. and Bursey, M. M.. 7. Org. Chem. 1966, 31, 1912-1917) Members of this class include diketoacid-containing analogues typified by L-731,988 (A) (Wai, J. S.; et al. 4-Aryl-2,4-dioxobutanoic acid inhibitors of HIV-I integrase and viral replication in cells. J. Med. Chem. 2000, 43, 4923-4926) and later-generation analogues such as the 7- carboxamido-8-hydroxy-l,6-naphthyridine L-870,812 (C) (Hazuda, D. J. et al Science 2004, 305, 528-532; Guare, J. P et al. Bioorg. Med. Chem. Lett. 2006, 16, 2900-2904) and the 5- hydroxy-6-oxo-4-pyrimidinecarboxamide clinical candidate from Merck, Raltegravir (MK0518, D; Crescenzi, B. et al.. PCT Application: WO 2003035077; Belyk, K. M. et al. PCT Application: WO2006060712; Markowitz, M. et al. J. Acquired Immun. Def. Syndromes 2006, 43, 509-515; Pace, P. et al. J. Med. Chem. 2007, 50, 2225-2239). The 3,4-dihydroxy- 2-pyridinecarboxamide E (Fuji, M. et al. JP 2004244320; Kong, L. C. C. et al. PCT Application: WO 2005042524) may be seen as a simplified variant of this latter structural class that retains key features needed for metal chelation.

[0004] Compound E bears similarity to the bis-salicylhydrazide G, which has previously been reported to inhibit ESf through metal chelation, but was effective only in assays using Mn²⁺ but not Mg²⁺ and lacked antiviral efficacy in HIV-I infected cells (Hong, H.et al. J. Med. Chem. 1997, 40, 930-936; Zhao, H.et al.. J. Med. Chem. 1997, 40, 937-941; Neamati, N. et al. J. Med. Chem. 1998, 41, (17), 3202-3209; Neamati, N. J. Med. Chem. 2002, 45, 5661-5670). Compound G differs from E both by being a hydrazide rather than a carboxamide and by containing a methylene at the 6-position of the aryl ring rather than a pyridyl nitrogen. Of greater significance in relation to potential metal chelating ability of G, is the absence of a second hydroxyl group at the salicyl 3-position that would correspond to the pyridyl 4-hydroxyl in E or the 6-oxo group in the 4-pyrimidinecarboxamide D.

[0005] Thus, there exists a clear need for identifying new and more effective inhibitors of retroviral integrases in order to provide the medical professionals with more effective means for the prevention and treatment of retroviral diseases. The present invention fulfills this and other related needs.

BRIEF SUMMARY OF THE INVENTION

[0006] The present invention relates to the identification of new inhibitors of retroviral integrase. This invention provides structural modifications of the original hydrazide 5a that could potentially affect metal chelating ability, and the preparation of new analogues generally represented by structure I (Figure 1). Thus, this invention provides a novel compounds and methods for inhibiting retrovirus proliferation. This method includes the step of contacting a cell, which is infected with a retrovirus or at risk of being infected with a retrovirus, with an effective amount of compound of formula I or II:

(I);

wherein R^ is H or -C_rC₈alkylaryl,

R² is H, OH or -C_rC_s alkoxy; each R^ is independently H or -Ci-Cg alkyl;

R⁴ is selected from the group consisting of: -C_rC₈alkylaryI, -C₃-C₂₄cycloalkyl, - alkylheteroaryl, -LN(R^4a)₂, or -LN=CH-aryl and -LN=CHheteraryl; each R^4a is independently selected from the group consisting of: H, -CO-aryl, - N=CH-aryl and -S0₂aryl; or is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(V); each R⁵ is independently selected from the group consisting of: haloC_rC₈alkyl-, -C_r Qalkylaryl, -halo, -amino, -NHSO₂R^5a, -N(SO₂R^5a)₂,, OH, -C,-C₈alkoxy; heteroC,-C₈alkyl- -COR⁵'¹ and -NHCOR⁵"; R^5a is aryl, OR^5b or NHR^5b; R^5b is -d-Cgalkyl or -NHN=CH-aryl;

R⁶ is -CO-aryl; -CH-aryl or -CH-C₃-C₂₄cycloalkyl;

Y¹ is CH₂, CO or SO₂;

Y² is CH₂, CO or SO₂; Y³ is N, CH or CR⁵;

L is a bond or a C2-C3alkenylene group; n is 1, 2 or 3; and each of each of aryl, heterocyclyl, heteraryl or cycloalkyl is optionally substituted with from one to five substituents independently selected from the group consisting of: C_rC₈alkyl, haloC,-C₈alkyl-, -C,-C₈alkylaryl, -halo, -amino, -NHSO₂R^5a, -N(SO₂R^5a)₂, OH, -C_rC₈alkoxy; heteroCpQalkyl-, heterocyclyl, -COC_rC₈alkyl, -CO₂C_rC₈alkyl; -COaryl and C0₂aryl; the dashed line indicates an optional bond; the wavy line indicates the point of attachment to the rest of the molecule; and pharmaceutically acceptable derivatives thereof.

[0007] In some embodiments, the compound has a formula:

In other embodiments, the compound has a formula:

(III). In some cases the compound has a formula:

[0008] In some embodiments of the above formulas, R* is H or -C_rC₈alkylaryl, R^ is H, OH or -

C_rC_g alkoxy; each R-* is independently H or -C_rC₈ alkyl; R⁴ is selected from the group consisting of: -C_rC₈alkylaryl, -C₃-C₂₄cycloalkyl, -alkylheteroaryl, -LN(R^4a)₂, or -LN=CH-aryl and - LN=CHheteraryl; each R^4a is independently selected from the group consisting of: H, -CO-aryl, - N=CH-aryl and -SO₂aryl; or is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(Va); each R⁵ is independently selected from the group consisting of: haloQ- Cgalkyl-, -C,-C₈alkylaryl, -halo, -amino, -NHSO₂R⁵³, -N(SO₂R^5a)₂, OH, -C_rC₈alkoxy; heterod-Csalkyl- -COR^5a and -NHC0R^5a; R^5a is Ci-Csalkyl, aryl, OR^5b or NHR^5b; R^5b is - Ci-Qalkyl or -NHN=CH-aryl; R⁶ is -CO-aryl; -CH-aryl or -CH-C₃-C₂₄cycloalkyl; Y¹ is CH₂, CO or SO₂; L is a bond or a C2-C3alkenylene group; n is 1, 2 or 3; and each of aryl, heterocyclyl, heteraryl or cycloalkyl is optionally substituted with from one to five substituents independently selected from the group consisting of: C,-C₈alkyl, haloC_rC₈alkyl-, -Ci-C₈alkylaryl, - halo, -amino, -NHSO₂R^5a, -N(SO₂R^5a)₂, OH, -C,-C₈alkoxy; heteroC,-C₈alkyl-, heterocyclyl, -COC,- Qalkyl, -CO₂C_rC₈alkyl; -COaryl and C0₂aryl; the dashed line indicates an optional bond; the wavy line indicates the point of attachment to the rest of the molecule; and pharmaceutically acceptable derivatives thereof. [0009] In yet other embodiments, the contacting step of the method is performed in vitro. In other embodiments, the cell is a part of a living animal, which may be a mammal such as a human, and the retrovirus may be a human retrovirus such as HIV-I . BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1: Structural features a general IN inhibitor (General structure i) common to diketo acid (A and B) and later generation inhibitors.

[0011] Figure 2: Synthesis scheme of hydrazides and amides with X as indicated in Table 1.

[0012] Figure 3: Synthesis scheme of bis-aroylhydrazines with R as indicated in Table 1.

[0013] Figure 4: Synthesis scheme of compounds wherein R and R¹ are as indicated in Table 1.

[0014] Figure 5: Synthesis scheme of compounds wherein X is as indicated in Table 2.

[0015] Figure 6: Synthesis scheme of compounds wherein R is as indicated in Table 3.

[0016] Figure 7: Synthesis scheme of compounds wherein R is as indicated in Table 3.

DETAILED DESCRIPTION OF THE INVENTION

DEFINITIONS

[0017] For the compounds of the invention, the term "alkyl," by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain, or cyclic hydrocarbon radical, or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include di- and multivalent radicals, having the number of carbon atoms designated (i.e., CpC₂₄ means one to twenty-four carbons). Examples of saturated hydrocarbon radicals include groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4- pentadienyl, 3-(l,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers. The term "alkyl," unless otherwise noted, is also meant to include those derivatives of alkyl defined in more detail below as "heteroalkyl." Alkyl groups which are limited to hydrocarbon groups are termed "homoalkyl". [0018] The term "alkylene" by itself or as part of another substituent means a divalent radical derived from an alkane, as exemplified by -CH₂CH₂CH₂CH₂-, and further includes those groups described below as "heteroalkylene." Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred in the present invention. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.

[0019] The terms "alkoxy," "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional sense, and refer to those alkyl groups attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively. [0020] The term "heteroalkyl," by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or cyclic hydrocarbon radical, or combinations thereof, consisting of the stated number of carbon atoms and from one to three heteroatoms selected from the group consisting of O, N, Si and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N and S may be placed at any interior position of the heteroalkyl group. The heteroatom Si may be placed at any position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule. Examples include -CH₂-CH₂-O-CH₃, -CH₂-CH₂-NH-CH₃, -CH₂-CH₂-N(CH₃)-CH₃, -CH₂-S- CH₂-CH₃, -CH₂-CH₂₁-S(O)-CH₃, -CH₂-CH₂-S(O)₂-CH₃, -CH=CH-O-CH₃, -Si(CH₃)₃, -CH₂- CH=N-OCH₃, and -CH=CH-N(CH₃)-CH₃. Up to two heteroatoms may be consecutive, such as, for example, -CH₂-NH-OCH₃ and -CH₂-O-Si(CH₃)₃. Similarly, the term "heteroalkylene" by itself or as part of another substituent means a divalent radical derived from heteroalkyl, as exemplified by -CH₂-CH₂-S-CH₂CH₂- and -CH₂-S-CH₂-CH₂-NH-CH₂-. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied.

[0021] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl", respectively. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include 1 -(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1 -piperazinyl, 2-piperazinyl, and the like. The terms "cycloalkylene" and "heterocycloalkylene" by themselves or as part of another substituent means a divalent radical derived from a cycloalkyl or heterocycloalkyl, respectively. [0022] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(Ci-C₄)alkyl" is mean to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like. [0023] The term "aryl" means, unless otherwise stated, a polyunsaturated, typically aromatic, hydrocarbon substituent which can be a single ring or multiple rings (up to three rings) which are fused together or linked covalently. The term "heteroaryl" refers to aryl groups (or rings) that contain from zero to four heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2- imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5- benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2- quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" by themselves or as part of another substituent means a divalent radical derived from an aryl or heteroaryl, respectively.

[0024] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above.

[0025] Each of the above terms (e.g., "alkyl," "heteroalkyl," "aryl" and "heteroaryl") are meant to include both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0026] Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be a variety of groups selected from: -OR', =0, =NR\ =N-OR', -NR'R", -SR', -halogen, -SiR¹R¹¹R¹", -OC(O)R', -C(O)R', -CO₂R', -CONR'R", -0C(0)NR'R", -NR¹¹C(O)R', -NR' -C(O)NR¹¹R¹", -NR¹¹C(O)₂R', -NH- C(NH₂)=NH, -NR' C(NH₂)=NH, -NH-C(NH₂)=NR' , -S(O)R', -S(O)₂R', -S(O)₂NR⁵R", -CN and -NO₂ in a number ranging from zero to (2m' +1), where m' is the total number of carbon atoms in such radical. R', R" and R'" each independently refer to hydrogen, unsubstituted (Ci-C₈)alkyl and heteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted alkyl, alkoxy or thioalkoxy groups, or aryHQ-C^alkyl groups. When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 5-, 6-, or 7-membered ring. For example, -NR'R'¹ is meant to include 1-pyrrolidinyl and 4-morpholinyl. From the above discussion of substituents, one of skill in the art will understand that the term "alkyl" is meant to include groups such as haloalkyl (e.g., -CF₃ and - CH₂CF₃) and acyl (e.g., -C(O)CH₃, -C(O)CF₃, -C(O)CH₂OCH₃, and the like).

[0027] Similarly, substituents for the aryl and heteroaryl groups are varied and are selected from: -halogen, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO₂, -CO₂R', -CONR'R", -

C(O)R', -OC(O)NR¹R", -NR¹¹C(O)R', -NR¹¹C(O)₂R', ,-NR' -C(O)NR¹¹R¹", -NH-C(NH₂)=NH, -NR'C(NH₂)=NH, -NH-C(NH₂)=NR\ -S(O)R', -S(O)₂R', -S(O)₂NR¹R", -N₃, -CH(Ph)₂, perfluoro(Ci-C₄)alkoxy, and perfluoro(Ci-C₄)alkyl, in a number ranging from zero to the total number of open valences on the aromatic ring system; and where R' , R" and R'" are independently selected from hydrogen, (Ci-C₈)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(Ci-C₄)alkyl, and (unsubstituted aryl)oxy-(Ci-C₄)alkyl.

[0028] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -T-C(O)-(CH₂)_q-U-, wherein T and U are independently -NH-, -0-, -CH₂- or a single bond, and q is an integer of from O to 2. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH₂)_r-B-, wherein A and B are independently -CH₂-, -0-, -NH-, -S-, -S(O)-, -S(O)₂-, -S(O)₂NR'- or a single bond, and r is an integer of from 1 to 3. One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -(CH₂)_S- X-(CH₂)_Γ, where s and t are independently integers of from O to 3, and X is -0-, -NR'-, -S-, - S(O)-, -S(O)₂-, or -S(O)₂NR'-. The substituent R' in -NR'- and -S(O)₂NR' - is selected from hydrogen or unsubstituted (C_rC₆)alkyl. [0029] As used herein, the term "heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).

[0030] The term "cycloalkylalkyl" refers to a cycloalkyl radical, as defined herein, attached to an alkyl radical, as defined herein. [0031] The term "alkoxy" refers to an alkyl ether radical containing from 1 to 24 carbon atoms. Exemplary alkoxyl groups include, but are not limited to, methoxyl, ethoxyl, n- propoxyl, /.rø-propoxyl, n-butoxyl, wo-butoxyl, sec-butoxyl, tert-butoxyl, and the like.

[0032] The term "heteroaryl" refers to aryl groups (or rings) that contain from one to five heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of heteroaryl groups include 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 1-pyrazolyl, 3-pyrazolyl, 2- imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4- isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3- thienyl, 2-ρyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, benzopyrazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5- quinoxalinyl, 3-quinolyl, 6-quinolyl and 1,2,3,4- tetra-hydroquinoline. Substituents for each of the above noted heteroaryl ring systems are selected from the group of acceptable substituents described above in aryl definition section. [0033] "Heterocyclyl" or "cycloheteroalkyl" means a saturated or unsaturated non-aromatic or aromatic cyclic or multicyclic radical of 3 to 24 ring atoms in which one or two ring atoms are heteroatoms selected from O, NR (where R is independently hydrogen or alkyl) or S(O)n (where n is an integer from 0 to 2), the remaining ring atoms being C, where one or two C atoms may optionally be replaced by a carbonyl group. The heterocyclyl ring may be optionally substituted independently with one, two, or three substituents selected from alkyl, cycloalkyl, cycloalkyl-alkyl, halo, nitro, cyano, hydroxy, alkoxy, amino, mono-alkylamino, di-alkylamino, haloalkyl, haloalkoxy, -COR (where R is hydrogen, alkyl, cycloalkyl, cycloalkyl-alkyl, phenyl or phenylalkyl), -(CR'R")_n-COOR (n is an integer from 0 to 5, R' and R" are independently hydrogen or alkyl, and R is hydrogen, alkyl, cycloalkyl, cycloalkyl- alkyl, phenyl or phenylalkyl), or -(CR'R")_n-CONRV (where n is an integer from 0 to 5, R' and R" are independently hydrogen or alkyl, R and R are, independently of each other, hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl). More specifically the term heterocyclyl includes, but is not limited to, tetrahydropyranyl, piperidino, N- methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl, 3-pyrrolidino, 2-pyrrolidon-l-yl, morpholino, thiomorpholino, thiomoφholino-1 -oxide, thiomorpholino- 1,1 -dioxide, pyrrolidinyl, and the derivatives thereof. The prefix indicating the number of carbon atoms (e.g., C3-C10) refers to the total number of carbon atoms in the portion of the cycloheteroalkyl or heterocyclyl group exclusive of the number of heteroatoms.

X Y X

[0034] "Heterocyclylalkyl" or "Cycloheteroalkyl-alkyl" means a radical -R R where R is y an alkylene group and R is a heterocyclyl group as defined herein, e.g., tetrahydropyran-2- ylmethyl, 4-methylpiperazin-l-ylethyl, 3-piperidinylmethyl, and the like. [0035] The term "amino" refers to RN(R)-, wherein R and R' are each independently a hydrogen atom, an alkyl group, an aryl group, a cycloalkyl, a heteroalkyl, a heterocycloalkyl or a heteroaryl, as defined herein.

[0036] The term "amide" refers to RC(O) N(R)-, wherein R and R' are each independently a hydrogen atom, an alkyl group, an aryl group, a cycloalkyl, a heteroalkyl, a heterocycloalkyl or a heteroaryl, as defined herein.

[0037] The term "carboxyl" refers to R(O)CO-, wherein R is a hydrogen atom, an alkyl group, a cycloalkyl, an aryl group, a heteroalkyl, a heterocycloalkyl or an heteroaryl ring, as defined herein.

[0038] The term "alkoxycarbonyl" refers to -C(O)OR, wherein R is an alkyl group, a cycloalkyl, a heteroalkyl, an arylalkyl, a heteroarylalkyl, as defined herein.

[0039] The term "arylalkyl" or "aralkyl" refers to an aryl radical, as defined herein, attached to an alkyl radical, as defined herein. Thus, the term "arylalkyl" or "aralkyl" is meant to include those radicals in which an aryl group is attached to an alkyl group. Non-limiting arylalkyl groups include benzyl, phenethyl, pyridylmethyl, 4-hydroxybenzyl, 3-fluorobenzyl, 2-fluorophenylethyl, and the like. Aralkyl groups also include those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced by, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(l-naphthyloxy)propyl, and the like).

[0040] The term "heteroaralkyl" means a radical -R^xR^y, where R^x is an alkylene group and R^y is a heteroaryl group as defined herein, e.g., ρyridine-3-ylmethyl, 3-(benzofuran-2- yl)propyl, and the like). [0041] The term "administration" or "administering" refers to various methods of contacting a substance with an animal, such as a mammal, especially a human. Modes of administration may include, but are not limited to, methods that involve contacting the substance intravenously, intraperitoneally, intranasally, transdermally, topically, subcutaneously, parentally, intramuscularly, orally, or systemically, and via injection, ingestion, inhalation, implantation, or adsorption by any other means. One exemplary means of administration of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention is via intravenous delivery, where the compound can be formulated as a pharmaceutical composition in the form suitable for intravenous injection, such as an aqueous solution, a suspension, or an emulsion, etc. Other means for delivering a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention includes intradermal injection, subcutaneous injection, intramuscular injection, or transdermal or transmucosal application as in the form of a cream, a patch, or a suppository.

[0042] An "effective amount" of a certain substance refers to an amount of the substance that is sufficient to effectuate a desired result. For instance, an effective amount of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention that is intended to inhibit the activity of an integrase of a retrovirus is an amount sufficient to achieve the goal of inhibiting the integrase when administered to a cell exposed to (or at risk of being exposed to) the retrovirus. The effect to be achieved may include the prevention, correction, or inhibition of progression of the symptoms of a disease/condition caused by infection by this retrovirus and related complications to any detectable extent. The exact quantity of an "effective amount" will depend on the purpose of the administration, and can be ascertainable by one skilled in the art using known techniques {see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); and Pickar, Dosage Calculations (1999)).

[0043] A "physiologically acceptable excipient" is an inert ingredient used in the formulation of a composition of this invention, which contains the active ingredient(s) of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention and is suitable for use, e.g., by injection into a patient in need thereof. This inert ingredient may be a substance that, when included in a composition of this invention, provides a desired pH, consistency, color, smell, or flavor of the composition.

[0044] The term "inhibit," "inhibiting," or "inhibition," when used in the context of how the activity of a retroviral integrase, e.g., HIV-I integrase, is affected, refers to any detectable negative change or decrease in quantity of a parameter that reflects the activity of a retroviral integrase, compared to a standard value. The level of this decrease, for example, in the activity of HIV-I integrase under a given condition following exposure to a hydrazide, amide, phthalimide and phthalhydrazide analog of the present invention from the same enzyme under the same condition not exposed to the compound or exposed to only a control compound having no known anti-integrase activity, is preferably at least 10% or 20%, and more preferably at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%, and most preferably 100%.

I. INTRODUCTION

[0045] Retroviruses are known to cause a variety of serious and even deadly diseases in humans. Most notably, the global HIV/ AIDS epidemic, caused by retrovirus human immunodeficiency virus (HIV), is expanding rapidly. The major priority for stopping the epidemic is developing new approaches to prevention, especially female-controlled measures including microbicides. Microbicides are medications formulated for vaginal administration to prevent the transmission of HIV and have several different mechanisms of action against HIV, such as 1) HIV inactivation by damaging viral structures (acid-buffering agents, surfactants etc.); 2) preventing HIV attachment to cells by interfering with CD4 receptor and CCR5 or CXCR4 co-receptors (negatively charged polymers such as cellulose sulphate, naphthalene sulphonate polymer, cellulose acetate 1,2-benzenedicarboxylate etc); and 3) prevention of HIV replication inside target cells (Stone and Jiang, Lancet 368(9534):431-433, 2006). Currently, all microbicides used in clinical trial to prevent HIV replication in the cell are reverse transcriptase inhibitors. However, inhibitors of reverse transcriptase could be toxic for human cells, due to the homology between reverse transcriptase and eukaryotic polymerases, and also could lead to the occurrence of drug resistant virus in case of prevention failure. Inhibitors of HIV integrase (IN) do not have these potential drawbacks because of its non-homology to mammalian enzymes (Marchand et al., Drug Discovery Today: Disease Mechanisms 3(253-260), 2006).

[0046] Like the integrase in every retrovirus, HIV IN is a key enzyme in the viral life cycle that inserts a DNA copy of the viral genome into the host DNA (Semenova et al. , Curr. Opin. in HIV/AIDS l(5):380-387, 2006). According to NMR data coupled with X-ray data, the protein consists of three domains: N-terminal, the core (or catalytic), and C-terminal. The N- terminal domain is thought to enhance EN multimerization that increases the concerted integration. The catalytic site of the enzyme includes three amino acids, Asp64, Aspl 16, and GIu 152 (D,D-35-E motif), necessary for the catalytic activity. These acidic residues coordinate at least one divalent cation (Mg²⁺ or Mn²⁺) that form a bridge with the DNA substrates. The C-terminal domain is responsible for metal-independent, nonspecific DNA binding through the amino acids 262 to 271. Previously, it was shown that in addition to this DNA-binding domain, the core domain of EN also contains a nonspecific but metal-dependent DNA-binding domain. No crystal structure of the whole protein IN or IN bound to DNA has been established. [0047] The insertion of a DNA copy of the viral genome into host DNA by HIV BSf proceeds by two reactions: first, the 3 '-processing reaction (3'-P), in which the proviral linear DNA synthesized by reverse transcription is hydrolyzed at the conserved CA dinucleotides from both of the 3 '-ends and the terminal pGpT dinucleotides are removed. The next stage, strand transfer (ST), proceeds in the nucleus through a transesterification reaction, where the processed 3'-OH of the viral cDNA is inserted into the backbone of the host DNA. A third IN-catalyzed reaction - disintegration, the reverse of ST, is found only in vitro and, unlike 3'- P and ST, could be catalyzed by core of IN alone.

[0048] Since the catalytic domain of HIV EN is required for enzyme functions, the development of inhibitors has been focused on affect on D,D-35-E motif with playing around different metal-affinity (Mg²⁺ vs. Mn²⁺) and different level of inhibition 3'-P vs. ST. Particular attention was given to the D,D-35-E motif after 5CITEP (a diketo acid-like derivative) was first co-crystallized in the catalytic domain of HEV IN and shown to bind in the D,D-35-E motif. Also, EN inhibitors currently in clinical trial have diketo-acid-like motifs that are believed to chelate divalent cations (Mg²⁺ or Mn²⁺) within the D,D-35-E motif and demonstrate preferential inhibition of the ST reaction. However, the removal part of protein (262-271 aa) or mutation in the positively charged amino acids (R-262, R-263 and K-264) within the metal-independent nonspecific DNA binding region (C-domain) has been shown to abolish 3'-P and ST activity, and to reduce (but not completely inhibit) disintegration activity (Lutzke et al, Nucleic Acids Res. 22(20) :4125-4131, 1994; and Nilsen et al, J. Virol. 70(3): 1580-1587, 1996). Thus, this region is believed to be essential for IN activity and can be a target for drug design.

[0049] The present inventors discovered that several hydrazide, amide, phthalimide and phthalhydrazide analogs (Tables), effectively blocks HIV IN activity in vitro and HIV replication in infected cell culture. Experiments comparing inhibition of wild-type IN to the IN core and antibodies mapping (probing inhibition of wild type DSf vs. core of IN coupled with antibodies mapping experiment) suggested that the most active hydrazide, amide, phthalimide and phthalhydrazide analogs bind outside the IN core and interferes with nonspecific DNA binding site in the C-domain. The high cytoprotective activity and novel binding domain makes the hydrazide, amide, phthalimide and phthalhydrazide analogs attractive for microbicides development.

II. SYNTHESIS OF HYDRAZIDE, AMIDE, PHTHALIMIDE AND PHTHALHYDRAZIDE ANALOG S

[0050] The present invention provides compounds of formula I or II:

(D;

wherein R* is H or -C_rC₈alkylaryl, R² is H, OH or -C , -C₈ alkoxy ; each R^ is independently H or -C,-C₈ alkyl;

R⁴ is selected from the group consisting of: -C_rC₈alkylaryl, -C₃-C₂₄cycloalkyl, - alkylheteroaryl, -LN(R^4a)₂, or -LN=CH-aryl and -LN=CHheteraryl; each R^4a is independently selected from the group consisting of: H, -CO-aryl, - N=CH-aryl and -SO₂aryl; or is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(V); each R⁵ is independently selected from the group consisting of: haloC_rC₈alkyl-, -C₁- Qalkylaryl, -halo, -amino, -NHSO₂R⁵¹¹, -N(SO₂R^5a)₂,, OH, -C,-C₈alkoxy; heteroC,-C₈alkyl- -COR^5a and -NHCOR^5a; R^5a is aryl, OR^5b or NHR^5b;

R^5b is -C-Cgalkyl or -NHN=CH-aryl;

R⁶ is -CO-aryl; -CH-aryl or -CH-C₃-C₂₄cycloalkyl;

Y' is CH₂, CO or SO₂;

Y² is CH₂, CO or SO₂; Y³ is N, CH or CR⁵;

L is a bond or a C2-C3alkenylene group; n is 1, 2 or 3; and each of aryl, heterocyclyl, heteraryl or cycloalkyl is optionally substituted with from one to five substituents independently selected from the group consisting of: Ci-C₈alkyl, haloC,- Qalkyl-, -C,-C₈alkylaryl, -halo, -amino, -NHSO₂R^5a, -N(SO₂R^5a)₂, OH, -C_rC₈alkoxy; heteroC,- Qalkyl-, heterocyclyl, -COQ-Qalkyl, -CO₂C,-C₈alkyl; -COaryl and C0₂aryl; the dashed line indicates an optional bond; the wavy line indicates the point of attachment to the rest of the molecule; and pharmaceutically acceptable derivatives thereof. [0051] In some embodiments, compounds have the formula Ha:

[0052] In other embodiments, compounds have the formula formula Ilia:

[0053] In some embodiments, compounds have the formula formula HIb:

[0054] In some embodiments, R¹ is H. In other embodiments, R¹ is -Ci-Qalkylaryl. [0055] In some embodiments, R^ is H. In other embodiments, R^ is OH. In other embodiments, R^ is -C, -C₈ alkoxy.

[0056] In some embodiments, R³ is H. In other embodiments, R³ is -C_rC₈ alkyl. [0057] In some embodiments, R⁴ is -C_rC_galkylaryl. In other embodiments, R⁴ is -C₃- C₂₄cycloalkyl. In other embodiments, R⁴ is -LN(R^4a)₂. In other embodiments, R⁴ is -LN=CH- aryl. In other embodiments, R⁴ is -LN=CHheteraryl. In other embodiments, R⁴ is -NHN=CH-Ph or -CH₂Ph. [0058] In some embodiments, R^4a has the formula VI:

wherein Y² is CH₂, CO or SO₂; and each Y³ is N, CH or CR⁵. In other embodiments, each R^4a or is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(Vb). [0059] In some embodiments, each R⁵ is independently selected from the group consisting of: - halo, -amino, -NHSO₂R^5a, -N(SO₂R⁵%, OH, -C,-C₈alkoxy, -COR^5a and -NHCOR^5a.

[0060] In some embodiments, Y¹ is CH₂. In other embodiments, Y¹ is CO. In other embodiments, Y¹ is SO₂. [0061] In some embodiments, L is a bond. In some embodiments, L is a C2-C3alkenylene group. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0062] In some embodiments, aryl is phenyl. In other embodiments, cycloalkyl is cyclopropyl. In other embodiments, C_rC₈ alkyl is -Me or Et. [0063] In some embodiments,the compounds are as described in the examples, tables and figures.

[0064] Various methods can be used to synthesize the compounds of formula I and II. A typical synthesis involves acylation and coupling or cyclization reactions. Other reactions can also be used for the synthesis of compound of formula I and II as shown in Figure 4. Figure 2 illustrates an exemplary general synthetic sequence.

[0065] As illustrated in Figure 2, starting from Corey's 2,3-dioxosulfinylbenzoylchloride (J), (Corey, E. J.; Bhattacharyya, S. Tetrahedron Lett. 1977, 45, 3919-392; Gramer, C. J.; Raymond, K. N. Org. Lett. 2001, 3, 2827-2830) the 2,3-dihydroxybenzoic acid hydrazides K and the dihydroxybenzamides L were prepared by reaction with the appropriate hydrazides or amines, respectively in dichloromethane at room temperature (see Figure 2; Pu, Y. et al. J. Org. Chem. 1994, 59,3642-3655).

[0066] The 2-hydroxy-3-methoxybenzoic acid hydrazides P were prepared by reaction of the pentafluorophenyl ester N with commercially-available hydrazides O in DMF at room temperature (see Figure 3A) (Zhao, H.; Burke, T. R., Jr. Tetrahedron 1997, 53, 4219-4230; (A. Basha et al., 1977; Zong et al., 2001) The benzoic acid hydrazides V, prepared by treating J and N with hydrazine, respectively, were converted to the hydrazones W by reaction with the corresponding aldehydes (see Figure 4).

[0067] Methylation of commercially- available 3,4-dimethoxybenzyl alcohol (X) (iodomethane / NaH in THF at 0 ⁰C) provided Y (93% yield), which was metalated (n- butyllithium) and quenched with methyl chloroformate to provide the methyl ester Z, AA (79% yield, Figure 5). (Napolitano, E. et al. J. Chem. Soc. Perkin 1 1986, 785-787.) Brief treatment with excess acetyl chloride in the presence of a catalytic amount of anhydrous zinc chloride directly produced the benzyl chloride BB, CC (86% yield) (Napolitano, E. et al. J. Chem. Soc. Perkin 11986, 785-787; Napolitano, E. et al. /. Org. Chem. 1983, 48, 3653- 3657). Synthesis of the series of JV-(l,3-dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)- benzamides (22a - p, Table 2) was readily achieved through a common methoxy-protected bicyclic hydrazide DD by acylation and final demethylation. Hydrazones 2Oo and 2Op were prepared by the reaction of hydrazide DD with aldehydes followed by demethylation as above. Refluxing with hydrazine in anhydrous acetonitrile afforded the key hydrazide DD (38% yield), which was acylated to afford the 2,3-dimethoxy-containing series EE. For the benzisothiazol hydrazide 21i, the acylating species required separate preparation. This consisted of metalating methyl ether Y (n-butyl lithium) followed by reaction with sulfuric chloride to give 5,6-dimethoxy-2-(methoxymethyl)benzene sulfonyl chloride. Treatment with anhydrous zinc chloride as described above for the preparation of BB, CC gave the corresponding benzyl chloride, which was reacted with hydrazide BB, CC to yield EE. Demethylation of the series EE (BBr₃ in dichloromethane) provided the final products FF. The series FF is characterized by a common bicyclic "right side" consisting of a 2,3- dihdroxy-substituted benzoylhydrazide conformationally restricted in which planarity was achieved by means of a ring-closing iV-methylene bridge.

[0068] Amides GG were synthesized from benzyl chloride BB, CC by refluxing with the appropriate amines in anhydrous acetonitrile, (Sahakitpichan, P.; Ruchirawat, S. Tetrahedron 2004, 60, 4169-4172; E. Harold Vickery, 1979) then demethylating as described above to yield the final products HH (see Figure 6).

[0069] The protected amides LL were synthesized from 3,4-dimethoxylphthalic anhydride (KK), which was obtained in three steps from 2,3-dimethoxyltoluene (Baudart, M. G.; Hennequin, L. F. J. Antibiotics 1993, 46, 1458-1470). Demethylation as described above gave the final products MM. Examples of protecting groups include those described and listed in Greene et al., Protective Groups in Organic Synthesis 3rd ed. Wiley, New York, 1999. III. ASSAY FOR RETROVIRAL INTEGRASE INHIBITORS

[0070] The functionality of the hydrazide, amide, phthalimide and phthalhydrazide analog s of this invention, i.e., their ability to inhibit the activity of a retroviral integrase (e.g., HIV-I integrase), can be tested and verified according to any one of the DNA binding assays or integration assays described in detail in this application or those known in the art. Once preliminarily identified, an inhibitor may be further tested in an in vitro activity assay in cultured cells, where the inhibitory effect of the compound is confirmed through a reduction in the cytopathic effect of a retrovirus on the cells.

A. DNA Binding Assay [0071] During the integration process, a retroviral integrase binds to viral DNA and catalyzes the insertion of viral DNA into the host cell DNA. Thus, the inhibitory capacity of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention can be tested in a DNA binding assay, in which the compound's ability to disrupt the interaction between a retroviral integrase and viral DNA is determined. [0072] Several methods are known to perform a DNA binding assay for this purpose. For instance, the Schiff-base assay can be used to screen for a potential HIV-I integrase inhibitor and is described in, e.g., Mazumder and Pommier, Nucleic Acids Res 23(15):2865-2871, 1995. Similar methods suitable for identifying inhibitors of other retrovirus integrase are described in, e.g., Terry et al, J. Virol., 62: 2358-2365, 1988; and Khan et al, Nucleic Acids Res., 19:851-860, 1990.

B. Disulfide Crosslinking Assay

[0073] A disulfide crosslinking assay is a second method for identifying an integrase inhibitor that exerts its effect by interfering with the binding between viral DNA and the integrase. This method is described in detail both in this application and Johnson et al, J. Biol. Chem. 281(l):461-467, 2006.

C. Catalytic Activity Assay

[0074] A catalytic activity or integration assay is another method for testing the inhibitory effect of a hydrazide, amide, phthalimide and phthalhydrazide analog on a retroviral integrase. In this assay format, the integrase activity or the inhibitory effect of candidate inhibitor is reflected by the level of viral DNA integration catalyzed by the integrase. An example of this method is described in full detail in the Example section of this disclosure for measuring HIV-I integrase activity, whereas similar methods suitable for measuring integrase activity of other retroviruses can be found in the literature, see, e.g., Fitzgerald et al, J. Virol., 66:6257-6263, 1992; Aiyar et al, J. Virol, 70:3571-3580, 1996; and Taganov et al, J. Virol, 78:5848-5855, 2004.

D. In vitro Activity Assay [0075] In addition, in vitro assays are available for confirming the inhibitory effect a candidate inhibitor of a retroviral integrase in cell culture, usually following a positive identification of the compound in the initial screening test such as the DNA binding assay or the integration assay. In this in vitro activity assay system, cells that are susceptible to infection by a particular type of retrovirus are first established in a stable culture. Under suitable conditions, the retrovirus is then introduced into the cultured cells, some of which also receive a pre-determined amount of a candidate inhibitor compound. Cell viability is then studied and the inhibitory effect of the compound is determined. The Example section of this application provides an example of such an assay system in which HIV- 1 integrase inhibitors were tested. A person of skill in the art would recognize, however, that a similar, cell culture-based system can be readily set up to confirm the function of a potential inhibitor of an integrase from another retrovirus.

IV. PHARMACEUTICAL COMPOSITION AND ADMINISTRATION

[0076] Another aspect of the present invention is a pharmaceutical composition comprising a hydrazide, amide, phthalimide and phthalhydrazide analog that is capable of inhibiting the activity of a retroviral integrase, e.g., HIV-I integrase. This composition, often further containing at least one physiologically acceptable excipient, can be used in anti-retroviral applications for both prophylactic and therapeutic purposes. Pharmaceutical compositions of the invention are suitable for use in a variety of drug delivery systems. Suitable formulations for use in the present invention are found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, 17th ed. (1985). For a brief review of methods for drug delivery, see, Langer, Science 249: 1527-1533 (1990).

[0077] For preparing pharmaceutical compositions containing a compound of the present invention, inert and pharmaceutically acceptable excipients or carriers are used. Liquid pharmaceutical compositions include, for example, solutions, suspensions, and emulsions suitable for intradermal, subcutaneous, parenteral, or intravenous administration. Sterile water solutions of the active component (e.g., a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention) or sterile solutions of the active component in solvents comprising water, buffered water, saline, PBS, ethanol, or propylene glycol are examples of liquid compositions suitable for parenteral administration. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, detergents, and the like.

[0078] Sterile solutions can be prepared by dissolving the active component (e.g., a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention) in the desired solvent system, and then passing the resulting solution through a membrane filter to sterilize it or, alternatively, by dissolving the sterile compound in a previously sterilized solvent under sterile conditions. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the preparations typically will be between 3 and 11, more preferably from 5 to 9, and most preferably from 7 to 8, or from 6.5 to 7. For instance, a 10 μM solution of NSC 34931 exhibits a pH of about 6.5-6.9 at room temperature.

[0079] In some embodiments, the compositions can be in solid or semi-solid formulations, using inert ingredients such as gelatin, ascorbate, trehalose, skim milk, starch, xylitol, and the like. [0080] The pharmaceutical compositions of the present invention can be administered by various routes, e.g., subcutaneous, intradermal, transdermal, intramuscular, intravenous, or intraperitoneal. In some cases, the composition is delivered by parenteral, intranasal, topical, oral, or local administration, such as by aerosol or transdermally, for prophylactic treatment. Frequently, the pharmaceutical compositions can be administered locally, e.g., deposited intra-vaginally or intra-rectally. Alternatively, the pharmaceutical compositions can be administered orally. Thus, the invention provides compositions for systemic, local, and oral administration, which comprise a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention dissolved or suspended in a physiologically acceptable carrier, preferably an aqueous carrier, e.g., water, buffered water, saline, PBS, and the like. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, detergents and the like. [0081] Alternatively, the composition can be delivered as a suppository or pessary. In some embodiments, the compound of this invention are prepared in a preservation matrix such as described in U.S. Pat. Nos. 6,468,526 and 6,372,209, and are delivered in a dissolvable element made of dissolvable polymer material and/or complex carbohydrate material selected for dissolving properties, such that it remains in substantially solid form before use, and dissolves due to human body temperatures and moisture during use to release the compound in a desired timed release and dosage. See, e.g., U.S. Pat. No. 5,529,782. The compound can also be delivered in a sponge delivery vehicle, such as described in U.S. Pat. No. 4,693,705, or via a tampon-like delivery tube. [0082] In some embodiments, the hydrazide, amide, phthalimide and phthalhydrazide analog is administered orally. For example, the physical form of the final recombinant products can be in a tablet/capsule suitable for oral ingestion, optionally in a sustained release formulation.

[0083] The preferred route of administering the pharmaceutical compositions is via intravenous injection at weekly dosage of about 1 μg - 10 mg, preferably 50 μg-1 mg, of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention for a 70 kg adult human. The appropriate dose may be delivered in daily, weekly, biweekly, or monthly intervals, by single or multiple administrations of the compositions with dose levels and pattern determined by the treating physician. In any event, the pharmaceutical formulations should provide a quantity of a hydrazide, amide, phthalimide and phthalhydrazide analog of this invention sufficient to effectively prevent or inhibit retroviral infection in an individual.

EXAMPLES

[0084] The following examples are provided by way of illustration only and not by way of limitation. Those of skill in the art will readily recognize a variety of non-critical parameters that could be changed or modified to yield essentially similar results.

General Synthetic Procedures for Compounds.

[0085] ¹H and ¹³C NMR data were obtained on a Varian 400 MHz spectrometer and are reported in ppm relative to TMS and referenced to the solvent in which the spectra were collected. IR spectra were run neat using a Jasco FT/IR-615 instrument. Solvent was removed by rotary evaporation under reduced pressure and anhydrous solvents were obtained commercially and used without further drying. Purification by silica gel chromatography was performed using EtOAc - hexanes solvent systems. Preparative high pressure liquid chromatography (HPLC) was conducted using a Waters Prep LC4000 system having photodiode array detection and using a YMC J 'sphere ODS-H80 column [YMC] (250 mm x 20 mm LD. ; S-4 μm particle size, 80 A pore) or a Phenomenex Ci₈ column [Phe] (250 mm x 21.2 mm LD. ; S-5 μm particle size, 110 A pore) at a flow rate of 10 mL/min. with binary solvent systems consisting of A = 0.1 % aqueous TFA and B = 0.1 % TFA in acetonitrile as indicated. Products were obtained as amorphous solids following lyophilization. High- resolution mass spectra (HRMS) were obtained from UCR Mass Spectrometry Facility, University of California at Riverside and fast-atom bombardment mass spectra (FABMS) were acquired with a VG Analytical 7070E mass spectrometer under the control of a VG 2035 data system.

2,3-Dihydroxybenzoic Acid Hydrazide (13a).

[0086] Hydrazide (or hydrazine, or diamino alkane) (1 mmol) was suspended in dichloromethane (1 mL) and 2,3-dioxosulfinylbenzoylchloride (7) (1 equivalent) (Corey, E. J. Tetrahedron Lett. 1977, 45, 3919-3922; Gramer, C. J.; Raymond, K. N. Org. Lett. 2001, 3, 2827-2830) (or 2 mmol for hydrazine and diamino alkane) followed by triethylamine (1 equivalent) (or 2 mmol for hydrazine and diamino alkane). The mixture stirred at room temperature (rt) overnight. The reaction was quenched by the addition of H₂O (0.5 mL) and the mixture was filtered to yield a solid product, which was further purified by HPLC using a preparative [YMC] column (linear gradient of 0% B to 50% B over 30 minutes; retention time = 13.0 minutes) was used to afford 13a as a white solid following lyophilization. ¹H NMR (DMSO) δ 7.21-7.18 (m, IH), 6.92-6.90 (m, IH), 6.70-6.65 (m, IH); ¹³C NMR (DMSO) <5 167.6, 148.1, 146.5, 119.4, 119.1, 118.4, 115.4; FAB-MS m/z 169 (M-H+).

Example 1 2-Hydroxybenzoic Acid 2-(2-Hydroxybenzoyl)hydrazide (5a).

[0087] Compound 5a was prepared as indicated in reference Zhao, H. et al. J. Med. Chem. 1997, 40, 937-941.

Examples 2-7 General Procedure A for the Synthesis of 2,3-Dihydroxybenzoic Acid Hydrazides 8a - 8e and 13.

[0088] To a suspension of hydrazide in dichloromethane was added 2,3- dioxosulfinylbenzoylchloride (7) (1 equivalent) (Corey, E. J. Tetrahedron Lett. 1977, 45, 3919-3922; Gramer, C. J.; Raymond, K. N. Org. Lett. 2001, 3, 2827-2830) followed by triethylamine (1 equivalent) and the mixture stirred at room temperature (overnight). The reaction was quenched by the addition of H₂O and the mixture was filtered to yield a solid product, which was purified by HPLC using a [YMC] column.

Example 2

2,3-Dihydroxybenzoic Acid 2-(2-Hydroxybenzoyl)hydrazide (8a).

[0089] Following General Procedure A, preparative HPLC with a linear gradient from 20% B to 40% B over 30 minutes; retention time = 22.4 minutes, provided 8a as a white solid following lyophilization. ¹H NMR (DMSO) δ 11.74 (bs, IH), 11.59 (bs, IH), 10.83 (bs, IH), 10.76 (bs, IH), 7.87 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.41 (dt, IH, J = 1.6 Hz, 7.2 Hz), 7.33 6.96- 6.90 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.96-6.90 (m, 3H), 6.72 (t, IH, J = 8.0 Hz); ¹³C NMR

(DMSO) δ 167.7, 166.8, 159.1, 148.9, 146.6, 134.6, 129.1, 119.7, 119.6, 119.1, 118.4, 117.7, 115.3, 114.9; FAB-MS m/z 287.1 (M-H). HRMS calcd for C₁₄Hi₃N₂O₅ [MH+]: 289.0824. Found: 289.0829.

Example 3

2,3-Dihydroxybenzoic Acid 2-(2,3-Dihydroxybenzoyl)hydrazide (8b).

[0090] Following General Procedure A, preparative HPLC with a linear gradient from 0% B to 50% B over 30 minutes; retention time = 23.6 minutes, provided 8b as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.64 (s, 2H), 10.77 (s, 2H), 9.38(bs, 2H), 7.33 (dd, 2H, / = 1.2 Hz, 8.0 Hz), 6.95 (dd, 2H, J = 1.2 Hz, 8.0 Hz), 6.72 (t, 2H, J = 8.0 Hz); ¹³C NMR (DMSO): δ 168.3 (2C), 149.1 (2C), 146.6 (2C), 119.8 (2C), 119.1 (2C), 118.0 (2C), 114.7 (2C); FAB-MS m/z 303.1 (M-H). HRMS calcd for Ci₄H₁₃N₂O₆ [MH+]: 305.0774. Found: 305.0779.

Example 4 2,3-Dihydroxybenzoic Acid 2-(2-Hydroxy-3-methoxybenzoyl)hydrazide (8c).

[0091] Following General Procedure A, preparative HPLC with a linear gradient from 5% B to 50% B over 30 minutes; retention time = 28.7 minutes, provided 8c as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.68 (s, IH), 11.61 (s, IH), 10.80 (d, 2H, J = 2.4 Hz), 9.40 (bs, IH), 7.46 (dd, IH, 7 = 1.6 Hz, 8.4 Hz), 7. 33 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.14 (d, IH, J = 8.0 Hz), 6.94 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.86 (t, IH, J = 8.4 Hz), 6.72 (t, IH, J = 8.0 Hz), 3.77 (s, 3H); FAB-MS m/z 317.1 (M-H).

Example 5

2,3-Dihydroxybenzoic Acid 2-(3-Hydroxy-2-naphthoyl)hydrazide (8d).

[0092] Following General Procedure A, preparative HPLC with a linear gradient from 5% B to 75% B over 35 minutes; retention time = 29.5 minutes, provided 8d as a white solid following lyophilization. ¹H NMR (DMSO): <5 11.61 (bs, IH), 11.38 (bs, IH), 10.97 (d, IH, J = 2.4 Hz), 10.91 (d, IH, J = 2.4 Hz), 9.43 (bs, IH), 8.50 (s, IH), 7.89 (d, IH, J = 8.4 Hz), 7.73 (d, IH, J = 8.4 Hz), 7.48 (t, IH, J = 8.0 Hz), 7.37 (d, IH, J = 7.6 Hz), 7.34 (t, IH, 7 = 8.0 Hz), 7.30 (s, IH), 6.95 (d, IH, J = 7.6 Hz), 6.73 (t, IH, 7 = 8.0 Hz); FAB-MS m/z 339.1 (MH⁺). HRMS calcd for C₁₈H₁₅N₂O₅ [MH+]: 339.0981. Found: 339.0995.

Example 6 2,3-Dihydroxybenzoic Acid 2-PicaIinoyIhydrazide (8e).

[0093] Following General Procedure A, preparative HPLC with a linear gradient from 5% B to 40% B over 35 minutes; retention time = 17.6 minutes, provided 8e as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.74 (bs, IH), 10.82 (s, IH), 10.71(s, IH), 9.03 (d, IH, 7 = 2.4 Hz), 8.75 (dd, IH, J = 1.6 Hz, 5.2 Hz), 8.24 (dt, IH, J = 2.4 Hz, 8.0 Hz), 7.55 (dd, IH, 7 = 4.8 Hz, 8.0 Hz), 7.33 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.95 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.72 (t, IH, 7 = 8.0 Hz); ¹³C NMR (DMSO): δ 189.0, 169.1, 164.7, 152.8, 149.4, 148.7, 146.7, 135.9, 128.5, 124.3, 119.8, 119.0, 114.6; FAB-MS m/z 274.1 (MH⁺). HRMS calcd for C_nH₁₂N₃O₄ [MH+]: 274.0828. Found: 274.0840.

Example 7

3-Hydroxy-pyridine-2-carboxylic acid N'-(2,3-dihydroxy-benzoyl)-hydrazide (8f).

[0094] Following General Procedure A, preparative HPLC with a linear gradient from 20% B to 45% B over 30 minutes; retention time = 19.1 minutes, provided 8f as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.78 (bs, IH), 1 1.06 (bs, IH), 10.72(s, IH), 8.19 (dd, IH, 7 = 1.2 Hz, 8.4 Hz), 7.44 (dd, IH, 7 = 1.2 Hz, 8.4 Hz), 7.33 (dd, IH, 7 = 1.2 Hz, 8.4 Hz), 6.95 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.72 (t, IH, 7 = 8.0 Hz); ¹³C NMR (DMSO): δ 168.9, 168.1, 157.6, 149.5, 146.7, 140.7, 130.9, 130.1 126.6, 1 19.9, 119.0, 118.0, 114.4; FAB-MS m/z 288.1 (M-H).

Example 8-10 General Procedure B for the Synthesis of iV-Alkyl-2,3-dihydroxybenzamides 9a - 9c.

[0095] To a solution of amine in dichloromethane was added 2,3- dioxosulfinylbenzoylchloride (7) (2 equivalents) followed by triethylamine (2 equivalents) and the mixture stirred at room temperature (overnight). The reaction was quenched by the addition of H₂O and the crude product was purified by HPLC using a [YMC] column.

Example 8

N,N'-l,3-Ethanediylbis[2,3-dihydroxybenzamide] (9a).

[0096] Following General Procedure B, preparative HPLC with a linear gradient from 10% B to 50% B over 30 minutes; retention time = 25.1 minutes, provided 9a as a white solid following lyophilization. ¹H NMR (CD₃OD): δ 8.66 (bs, 2H), 7.16 (dd, 2H, J = 1.6 Hz, 8.0 Hz), 6.88 (dd, 2H, 7= 1.6 Hz, 8.0 Hz), 6.67 (t, 2H, J = 8.0 Hz), 3.59-3.58 (m, 4H); ¹³C NMR (CDCl₃): δ 170.7 (2C), 148.9(2C), 145.9(2C), 118.2(2C), 118.1(2C), 117.2(2C), 115.2(2C), 39.0, 38.8; FAB-MS m/z 333.3 (MH⁺). HRMS calcd for C₁₆H₁₇N₂O₆ [MH+]: 333.1087. Found: 333.1090.

Example 9 N,N' - 1,3-Propanediy lbis[2,3-dihydroxybenzamide] (9b).

[0097] Following General Procedure B, preparative HPLC with a linear gradient from 10% B to 55% B over 30 minutes; retention time = 24.7 minutes, provided 9b as a white solid following lyophilization. ¹U NMR (DMSO): δ 8.75 (t, 2H, J = 10.8 Hz), 7.21 (dd, 2H, J = 1.6 Hz, 8.0 Hz), 6.86 (dd, 2H, J = 1.6 Hz, 8.0 Hz), 6.63 (dt, 2H, J = 1.6 Hz, 8.0 Hz), 3.30 (dd, 4H, J = 6.8 Hz, 13.2 Hz), 1.77 (dt, 2H, J = 6.8 Hz, 13.2 Hz); FAB-MS m/z 345.5 (M-H⁺). HRMS calcd for C_nH₁₉N₂O₆ [MH+]: 347.1243. Found: 347.1247. Example 10 N-(4-Fluorobenzyl)-2,3-dihydroxybenzamide (9c).

[0098] Following General Procedure B, the reaction was run initially at -20 ⁰C then stirred at room temperature (overnight). Preparative HPLC with a linear gradient from 40% B to 100% B over 35 minutes; retention time = 21.5 minutes, provided 9c as a white solid following lyophilization. ¹H NMR (DMSO): δ 12.55 (bs, IH), 9.28 (t, IH, J = 6.0 Hz), 9.13 (bs, IH), 7.34-7.27 (m, 3H), 7.13-7.09 (m, 2H), 6.88 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.65 (t, IH, J = 8.0 Hz), 4.43 (d, 2H, J = 6.0 Hz); ¹³C NMR (DMSO): δ 170.1, 162.9, 160.5, 150.1, 146.7, 135.6 (d, 1C, J = 3.0 Hz), 129.8 (d, 1C, J = 7.7 Hz), 119.4, 118.5, 1 17.7, 115.6, 115.4, 115.3, 42.1 ; FAB-MS m/z 262.1 (MH⁺). HRMS calcd for C₁₄H₁₃FNO₃ [MH⁺]: 262.0879. Found: 262.0872.

Examples 11-14

General Procedure C for the Synthesis of 2-Hydroxy-3-methoxybenzoic Acid Hydrazides 12a - 12c and 13b. [0099] A solution of the appropriate 2-aroylhydrazide (for products 12a - c) or hydrazine (for product 13b) (1 equivalent) and 3-methoxysalylic acid pentafluorophenyl ester (10, 1 equivalent), prepared in a fashion similar to that reported for salicylic acid pentafluorophenyl ester, (Zhao, H. et al. Tetrahedron 1997, 53, 4219-4230) in DMF was stirred at room temperature (overnight). Crude product was collected by filtration and purified by preparative [YMC] HPLC.

Example 11

2-Hydroxy-3-methoxybenzoic Acid 2-(2-Hydroxy-3-methoxybenzoyl)hydrazide (12a).

[0100] Following General Procedure C, preparative HPLC with a linear gradient from 5% B to 70% B over 35 minutes; retention time = 27.4 minutes, provided 12a as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.66 (s, 2H), 10.82 (s, 2H), 7.45 (dd, 2H, J = 1.2 Hz, 8.0 Hz), 7. 14 (dd, 2H, J = 1.2 Hz, 8.0 Hz), 6.86 (t, 2H, J = 8.0 Hz), 3.78 (s, 6H); FAB-MS m/z 333.1 (MH⁺). HRMS calcd for C₁₆H_nN₂O₆ [MH+]: 333.1087. Found: 333.1099. Example 12

2-Hydroxy-3-methoxybenzoic Acid 2-(3-Hydroxy-2-naphthoyl)hydrazide (12b).

[0101] Following General Procedure C, preparative HPLC with a linear gradient from 30% B to 70% B over 30 minutes; retention time = 22.3 minutes, provided 12b as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.67 (bs, IH), 11.42 (bs, IH), 11.01 (bs, 2H), 8.51 (s, IH), 7.88 (d, IH, J = 8.0 Hz), 7.32 (d, IH, / = 8.0 Hz), 7.51-7.45 (m, 2H), 7.34-7.30 (m, 2H), 7.13 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.87 (t, IH, J = 8.0 Hz), 3.78 (s, 3H); ¹³C NMR (DMSO): δ 166.6, 165.1, 154.1, 149.5, 148.8, 136.4, 131.4, 131.3, 129.3, 127.2, 126.3, 124.3, 120.0, 119.9, 119.2, 116.2, 115.2, 111.1, 56.4; FAB-MS m/z 353.1 (MH⁺). HRMS calcd for C₁₉H₁₇N₂O₅ [MH+]: 353.1137. Found: 353.1134.

Example 13 2-Hydroxy-3-methoxybenzoic Acid 2-Picalinoylhydrazide (12c).

[0102] Following General Procedure C, preparative HPLC with a linear gradient from 5% B to 40% B over 35 minutes; retention time = 18.4 minutes, provided 12c as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.79 (bs, IH), 10.85 (s, IH), 10.74(bs, IH), 9.03 (d, IH, J = 1.6 Hz), 8.73 (dd, IH, J = 1.6 Hz, 4.8 Hz), 8.22 (dt, IH, J = 2.0 Hz, 8.0 Hz), 7.54-7.51 (m, IH), 7.46 (dd, IH, J = 1.2 Hz, 8.0 Hz), 7.13 (dd, IH, J = 1.2 Hz, 8.0 Hz), 6.85 (t, IH, 7 = 8.0 Hz), 3.77 (s, 3H); ¹³C NMR (DMSO): δ 168.6, 164.7, 153.1, 150.3, 148.9, 135.7, 128.4, 124.1, 119.6, 1 19.4, 119.0, 116.3, 114.7, 56.4; FAB-MS m/z 288.1 (MH⁺). HRMS calcd for C₁₄H₁₄N₃O₄ [MH+]: 288.0984. Found: 288.0984. Example 14 2-Hydroxy-3-methoxybenzoic Acid Hydrazide (13b).

[0103] Following preparation according to General Method C, purification by preparative HPLC using a [Phe] column with a linear gradient from 5% B to 20% B over 35 minutes; retention time = 18.6 minutes, provided 13b as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.31 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.11 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.82 (t, IH, J = 8.0 Hz), 3.78 (s, 3H); ¹³C NMR (DMSO): δ 166.9, 148.7, 148.5, 120.2, 119.3, 116.1, 115.6, 56.5; FAB-MS m/z 182.1 (MH⁺). HRMS calcd for C₈H₁₁N₂O₃ [MH+]: 183.0770. Found: 183.0773.

Examples 15-18 General Procedure D for the Synthesis of Hydrazones 14a - 14c.

[0104] Hydrazide 13a (for products 14a and 14b) or 13b (for product 14c) (1 mmol) was suspended together with the appropriate aldehyde (1 mmol) in THF (2 mL) and the mixture was stirred at room temperature (overnight). The crude product was collected by filtration and purified directly by HPLC using a [YMC] column.

Example 15

2,3-Dihydroxybenzoic Acid [(2,3-Dihydroxyphenyl)methylene]hydrazide (14a).

[0105] Following preparation according to General Method D, purification by preparative HPLC with a linear gradient from 20% B to 50% B over 35 minutes; retention time = 27.0 minutes, provided 14a as a white solid following lyophilization. ¹H NMR (DMSO): δ 8.49 (s, IH), 7.24 (d, IH, J = 8.0 Hz), 6.93 (dt, 2H, J = 1.6 Hz, 8.0 Hz), 6.82 (dd, IH, / = 1.6 Hz, 8.0 Hz), 6.73 (dd, 2H, J = 8.0 Hz, 16.8 Hz); ¹³C NMR (DMSO): δ 165.6, 150.7, 148.2, 146.0, 145.4, 121.2, 120.0, 119.6, 119.7, 118.8 (2C), 118.5, 118.1, 115.5; FAB-MS m/z 287.1 (M- H). HRMS calcd for Ci₄Hi₃N₂O₅ [MH+]: 289.0824. Found: 289.0825. Example 16

2,3-Dihydroxybenzoic Acid [(2,3-Dimethoxyphenyl)methylene]hydrazide (14b).

[0106] Following preparation according to General Method D, purification by preparative HPLC with a linear gradient from 5% B to 60% B over 35 minutes; retention time = 28.6 minutes, provided 14b as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.92 (s, IH), 11.86 (s, IH), 8.42 (s, IH), 7.43 (dd, IH, 7 = 3.2 Hz, 6.0 Hz), 7.33 (dd, IH, J = 1.2 Hz, 8.4 Hz), 7.11-7.09 (m, 2H), 6.93 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.72 (t, IH, J = 8.0 Hz), 3.79 (s, 3H), 3.76 (s, 3H); ¹³C NMR (DMSO): δ 166.5, 153.1, 149.7, 148.6, 146.7, 145.0, 127.9, 124.8, 119.6, 118.8, 118.0, 117.5, 115.4, 114.9, 61.7, 56.2; FAB-MS m/z 315.2 (M-H). HRMS calcd for C₁₆H₁₇N₂O₅ [MH+]: 317.1137. Found: 317.1144.

Example 17

2-[(2,3-Dihydroxy-benzoyl)-hydrazonomethyl]-cyclopropanecarboxylic acid ethyl ester (14d).

[0107] Following preparation according to General Method D, purification by preparative HPLC with a linear gradient from 5% B to 55% B over 35 minutes; retention time = 25.3 minutes, provided 14d as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.55 (s, IH), 7.40 (d, IH, J = 7.2 Hz), 7.21 (d, IH, 7 = 8.0 Hz), 6.90 (d, IH, J = 8.0 Hz), 6.67 (t, IH, /= 8.0 Hz), 4.06 (dd, 2H, / = 7.2 Hz, , J = 14.4 Hz), 2.09-2.01 (m, 2H), 1.37-1.28 (m, 2H), 1.16 (dd, 3H, , J = 7.2 Hz, , 7 = 11.9 Hz); ¹³C NMR (DMSO): δ 199.7, 172.1, 165.8, 152.6, 149.4, 146.6, 119.5, 118.7, 115.5, 60.9; FAB-MS m/z 293 (MH+).

Example 18

2-Hydroxy-3-methoxybenzoic Acid [(4-Fluorophenyl)methylene]hydrazide (14c).

[0108] Following preparation according to General Method D, purification by preparative HPLC with a linear gradient from 45% B to 55% B over 30 minutes; retention time = 15.4 minutes, provided 14c as a white solid following lyophilization. ¹H NMR (CDCl₃): δ 10.80 (s, IH), 10.27 (s, IH), 8.24 (s, IH), 7.63-7.60 (m, 2H), 7.52 (d, IH, J = 8.0 Hz), 6.98-6.93 (m, 2H), 6.80 (t, IH, 7 = 8.0 Hz), 3.85 (s, IH); ¹³C NMR (CDCl₃): δ 165.3, 165.1, 162.8, 149.0, 148.1, 129.7 (d, 1C, J = 3.0 Hz), 129.6, 129.5, 119.7, 119.2, 115.9, 115.7, 115.0, 114.9, 59.2; FAB-MS m/z 289.1 (MH⁺). HRMS calcd for C₁₅H₁₄FN₂O₃ [MH+]: 289.0988. Found: 289.0991.

Example 19 iV-(4-FIuorobenzyl)-2-hydroxy-3-methoxybenzamide (15).

[0109] To a solution 3-methoxysalicylic acid pentfluorophenyl ester (10) (479 mg, 1.43 mmol) in anhydrous dichloromethane (10 mL) was added 4-fluorobenzlamine (0.25 mL, 2.20 mmol) and the solution was stirred at room temperature (overnight). The mixture was concentrated and purified by silica gel column chromatography to provide the crude product as a solid (361 mg, 92% crude yield). Further purification was achieved by preparative HPLC [YMC] with a linear gradient from 40% B to 55% B over 30 minutes; retention time = 25.6 minutes and yielded 15 as a white solid following lyophilization.¹H NMR (CDCl₃): δ 7.26- 7.21 (m, 3H), 7.12 (dd, IH, / = 1.6 Hz, 8.0 Hz), 6.97-6.91 (m, 2H), 6.74 (dd, IH, J = 6.4 Hz, 8.0 Hz), 4.53 (d, 2H, J = 6.0 Hz), 3.82 (s, 3H); ¹³C NMR (CDCl₃): δ 169.2, 163.4, 161.0,

150.5, 148.7, 133.5 (d, 1C, J = 3. I Hz), 129.5 (d, 1C, J = 8.4 Hz), 118.4, 118.0, 115.6, 115.4, 115.0, 114.7, 56.1, 42.9; FAB-MS m/z 21 Aλ (M-H). HRMS calcd for C₁₅H₁₅FNO₃ [MH+]: 276.1036. Found: 276.1037.

Example 20 l,2-Dimethoxy-3-(methoxymethyl)benzene (17).

[0110] Sodium hydride (95% in oil, 5.22 g, 0.207 mol) was added portion-wise to a solution of 3,4-dimethoxybenzyl alcohol (16) (23 mL, 0.158 mol) in anhydrous THF (150 mL) at 0 ⁰C and the resulting mixture was stirred at 0 ⁰C (10 minutes). To this was added iodomethane (12.86 mL, 0.207 mol) drop-wise and the mixture was allowed to come to ambient temperature and stirred (3 h). The reaction was quenched by the addition of ice and EtOAc, extracted with EtOAc and the combined organic phase was washed with brine and dried (Na₂SO₄). Evaporation of solvent provided 17 as a colorless residue (27.8 g, 93% yield). ¹H NMR (CDCl₃): δ 6.83-6.77 (m, 2H), 4.32 (s, 2H), 3.82 (s, 3H), 3.80 (s, 3H), 3.30 (s, 3H); ¹³C NMR (CDCl₃): δ 149.0, 148.5, 130.7, 120.2, 110.9, 110.8, 74.5, 57.8, 55.8, 55.7; FAB-MS m/z 182.1 (M⁺).

Example 21

2,3-Dimethoxy-6-(methoxymethyl)benzoic Acid Methyl Ester (18).

[0111] To a solution of methyl ether 17 (1.Og, 5.49 mmol) in anhydrous ether (15 mL) was added n-butyl lithium (1.6 M in hexanes, 6.4 mmol) dropwise with stirring at 0 ⁰C (1 h). The resulting precipitate suspension was cooled to -80 ⁰C and methyl chloroformate (2.0 mL, 26.0 mmol) was added then the reaction mixture was allowed to return to room temperature. The mixture was partitioned between H₂O and ether and the organic phase was dried (Na₂SO₄) and concentrated to provide a residue, which was purified by silica gel column chromatography to yield 18²⁴ (24. Napolitano, E.; Spinelli, G.; Fiaschi, R.; Marsili, A. A smple total synthesis of the isoindolobenzazepine alkaloids lennoxamine and chilenamine. /. Chem. Soc. Perkin 11986, 785-787.) as a colorless oil (1.04 g, 79% yield). ¹H NMR (CDCl₃): δ 6.96-6.93 (m, IH), 6.83-6.81 (m, IH), 4.31 (s, 2H), 3.82 (s, 3H), 3.77 (s, 3H), 3.23 (s, 3H); ¹³C NMR (CDCl₃): δ 167.7, 152.2, 146.4, 128.2 (2C), 124.2, 113.1, 72.0, 61.4, 58.0, 55.8, 52.0; FAB-MS m/z 240.1 (M⁺).

Example 22

6-(Chloromethyl)-2,3-dimethoxybenzoic Acid Methyl Ester (19).

[0112] Acetyl chloride (0.36 mL, 5.07 mmol) was added drop-wise with stirring to a solution of methyl ether 18 (362 mg, 1.51 mmol) and anhydrous zinc chloride (10 mg, 0.07 mmol) in anhydrous ether (3 mL) at 0 ⁰C. After 30 minutes aluminum oxide (360 mg) was added and the mixture filtered through a short pad of aluminum oxide. The eluent was evaporated and the residue was purified by silica gel column chromatography to yield 19 (Napolitano, E. et al. J. Chem. Soc. Perkin 11986, 785-787; Napolitano, E. et al. J. Chem. Soc, Perkin Trans 1 1987, 12, 2565-2568) as colorless oil (317 mg, 85.9% yield).¹H NMR (CDCl₃): δ 7.01 (d, IH, / = 8.4 Hz), 6.82 (d, IH, J = 8.4 Hz), 4.51 (s, 2H), 3.86 (s, 3H), 3.79 (s, 3H), 3.77 (s, 3H); ¹³C NMR (CDCl₃): δ 167.1, 153.0, 146.7, 128.6, 127.3, 125.7, 113.4, 61.4, 55.8, 52.4, 43.6; FAB-MS m/z 245 (MH⁺).

Example 23

2-Amino-2,3-dihydro-7,8-dimethoxy-lH-isoindol-l-one (20).

[0113] Anhydrous hydrazine (0.44 mL, 14.1 mmol) was added to a solution of benzyl chloride 19 (3.45 g, 14.1 mmol) in anhydrous acetonitrile (10 mL) and the solution was stirred at reflux (3 h). Solvent was removed under reduced pressure, the residue was partitioned between H₂O and methylene chloride and the organic phase was dried (Na₂SO₄) and evaporated to dryness to provide a residue, which was crystallized from EtOAc - hexanes (20 : 1) to provide 20 as a white solid (1.11 g, 38% yield). ¹H NMR (CDCl₃): δ 7.02 (dd, 2H, J = 8.0 Hz, 14.4 Hz), 4.36 (s, 2H), 4.28 (bs, 2H), 4.02 (s, 3H), 3.84 (s, 3H); ¹³C

NMR (CDCl₃): δ 166.1, 152.2, 147.0, 132.6, 123.5, 117.8, 116.3, 62.5, 56.6, 52.0; FAB-MS m/z 209.1 (MH⁺).

Examples 24-30 General Procedure E for the Synthesis of Hydrazides 21a - 21h. [0114] Triethylamine ( 1.0 mmol) was added drop-wise to a solution of hydrazide 20 ( 1.0 mmol) and the appropriate acid chloride (1.0 mmol) or pentafluorophenyl ester (1.0 mmol) or anhydride (0.5 mmol) in anhydrous dichloromethane (2.0 mL) and the reaction mixture was stirred at room temperature (overnight). The mixture was partitioned between brine - EtOAc and the combined organic phase was dried (Na₂SO₄) and evaporated to dryness and residue was purified by silica gel column chromatography.

Example 24

Λ/-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-benzamide (21a).

[0115] Following general procedure E using benzoic anhydride as an acylating reagent provided product 21a in 90% yield. ¹H NMR (CDCl₃): δ 10.42 (s, IH), 7.78 (dd, 2H, J = 0.1 Hz, 8.4 Hz), 7.32 (dt, IH, J = 0.8 Hz, 8.0 Hz), 7.18 (t, 2H, J = 8.4 Hz), 7.02 (d, IH, J = 8.0 Hz), 6.95 (d, IH, J = 8.0 Hz), 4.54 (s, 2H), 3.96 (s, 3H), 3.80 (s, 3H); ¹³C NMR (CDCl₃): δ 166.9, 166.3, 152.0, 147.4, 133.7, 132.0, 130.9, 129.9, 128.3, 128.2, 127.6, 122.4, 118.1, 117.3, 62.4, 56.6, 50.5; FAB-MS m/z 313.1 (MH⁺).

Example 25 N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-(2-hydroxy)benzamide (21b).

[0116] Following general procedure E using salicylic acid pentafluorophenyl ester as an acylating reagent provided product 21b in 59% yield. ¹H NMR (CDCl₃): «5 11.48 (s, IH), 10.82 (s, IH), 7.77 (dd, IH, J = 1.2 Hz, 8.0 Hz), 7.19 (dt, IH, J = 1.2 Hz, 8.4 Hz), 7.06 (d, IH, J = 8.4 Hz), 6.99 (d, IH, J = 8.4 Hz), 6.69-6.64 (m, 2H), 4.52 (s, 2H), 3.95 (s, 3H), 3.80 (s, 3H); ¹³C NMR (CDCl₃): δ 169.3, 167.0, 160.8, 152.1, 147.5, 134.6, 133.5, 127.1, 122.2, 118.9, 118.2, 117.9, 117.6, 112.4, 62.3, 56.6, 50.6. FAB-MS m/z 327.1 (M-H).

Example 26

7V-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-(2,3-dimethoxy)benzamide (21c).

[0117] Following general procedure E using 2,3-dimothoxybenoyl chloride as an acylating reagent provided product 21c in 91% yield. ¹H NMR (CDCl₃): δ 9.96 (s, IH), 7.55 (dd, IH, 7 = 2.0 Hz, 8.0 Hz), 7.05-6.97 (m, 4H), 4.53 (s, 2H), 3.99 (s, 3H), 3.96 (s, 3H), 3.79 (s, 3H), 3.78 (s, 3H); ¹³C NMR (CDCl₃): δ 165.4, 165.1, 152.6, 152.0, 147.8, 147.4, 133.5, 124.6, 124.5, 122.7, 122.6, 118.1, 117.0, 116.2, 62.3, 62.0, 56.5, 56.0, 50.6; FAB-MS m/z 373 (MH⁺).

Example 27

N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-iV-(4-fIuorobenzoyI)-4- fluorobenzamide (2Id).

[0118] Following general procedure E using 4-fluorobenoyl chloride as an acylating reagent provided the diacylated product 21d in 57% yield. ¹H NMR (CDCl₃): δ 7.88-7.84 (m, 4H), 7.12 (d, IH, 7 = 8.0 Hz), 7.07-7.03 (m, 4H), 6.99 (d, IH, 7 = 8.0 Hz), 4.48 (s, 2H), 4.04 (s, 3H), 3.85 (s, 3H); ¹³C NMR (CDCl₃): δ 170.5 (2C), 166.7, 165.8, 164.2, 152.4, 148.1, 133.5 (d, 2C, 7 = 9.1 Hz), 133.3, 131.4 (2C), 131.3 (2C), 129.3, 129.5, 118.2, 118.1, 116.0, 115.9 (d, 2C, 7 = 22.1 Hz), 62.5, 56.7, 50.6; FAB-MS m/z 453.1 (MH⁺).

Example 28

Λ^r-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-N-(4-fluorobenzenesulfonyI)-4- flourobenzesulfonamide (2If).

[0119] Following general procedure E using 4-fluorobenzenesulfonyl chloride as an acylating reagent provided the diacylated product 21f in 52% yield. ¹H NMR (CDCl₃): δ 7.93-7.90 (m, 4H), 7.17-7.12 (m, 5H), 7.05 (dd, IH, 7 = 0.4 Hz, 8.0 Hz), 4.69 (s, 2H), 3.89 (s, 3H), 3.85 (s, 3H); ¹³C NMR (CDCl₃): δ 167.5, 165.0, 164.9, 152.3, 148.1, 133.7 (d, 2C, 7 = 3.0 Hz), 133.3, 132.3 (2C), 132.2 (2C), 120.4, 118.5, 118.3, 116.5 (2C), 116.2 (2C), 62.3, 56.7, 51.8; FAB-MS m/z 525 (M-H).

Example 29 iV-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoiπdoI-2-yl)-N-(3,4- dimethoxybenzenesulfonyl)-3,4-dimethoxybenzesulfonamide (2Ig).

[0120] Following general procedure E using 3,4-dimethoxybenzenesulfonyl chloride as an acylating reagent provided the diacylated product 21g in 43% yield.Η NMR (CDCl₃): δ 7.51 (dd, 2H, / = 1.2 Hz, 8.4 Hz), 7.41 (d, 2H, J = 1.6 Hz), 7.13 (d, IH, J = 8.0 Hz), 7.02 (d, IH, 7 = 8.0 Hz), 6.84 (d, 2H, J = 8.4 Hz), 4.65 (s, 2H), 3.89 (s, 9H), 3.83 (s, 3H), 3.80 (s, 6H); ¹³C NMR (CDCl₃): δ 164.9, 154.0 (2C), 152.2, 148.8 (2C), 148.0, 133.5, 129.3 (2C), 123.7 (2C), 120.8, 118.2 (2C), 111.4 (2C), 110.0 (2C), 62.3 (2C), 56.7 (2C), 56.2 (2C), 51.8; FAB-MS m/z 609.1 (MH⁺).

Example 30 6,6^t,7,7'-Tetramethoxy-[2,2'-bi-2H-isoindole]-l,l'(3H,3Η)-dione (21h).

[0121] Following general procedure E using benzyl chloride 19 as an acylating reagent provided product 21h in 41% yield following crystallization from EtOAc - hexanes (10 : 1). ¹K NMR (CDCl₃): δ 7.12 (d, 2H, J = 8.0 Hz), 7.06 (d, 2H, J = 8.0 Hz), 4.62 (s, 4H), 4.03 (s, 6H), 3.86 (s, 6H); ¹³C NMR (CDCl₃): δ 165.5 (2C), 152.2 (2C), 147.7 (2C), 133.3 (2C), 122.5 (2C), 118.2 (2C), 117.4 (2C), 62.5 (2C), 56.7(2C), 49.0(2C); FAB-MS m/z 385.1 (MH⁺).

Example 31

N-[(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-N'-[(6,7-dimethoxy-l,l-dioxido- l,2-benzisothiazol-2(3H)-yl)] Hydrazide (2Ii).

[0122] To a solution of methyl ether 17 (857 mg, 4.71 mmol) was added drop-wise rc-butyl lithium (1.6 M in hexane, 3.24 mL, 5.18 mmol) at O ⁰C. The resulting white suspension was stirred at O ⁰C (1 h), then cooled (-78 ⁰C) and sulfuric chloride (1.2 mL, 14.8 mmol) was added, then the mixture was allowed to come to room temperature and stirred (overnight). The reaction was quenched by the addition of MeOH (1.0 mL) while stirring (2 h). Solvent was removed by evaporation and residue purified by silica gel column chromatography to afford intermediate 5,6-dimethoxy-2-(methoxymethyl)benzenesulfonyl chloride as a colorless oil (737 mg, 55.7% yield) [¹H NMR (CDCl₃): δ 7.35 (d, IH, 7 = 8.8 Hz), 6.18 (d, IH, 7 = 8.8 Hz), 4.71 (s, 2H), 3.99 (s, 3H), 3.88 (s, 3H), 3.40 (s, 3H); ¹³C NMR (CDCl₃): δ 153.0, 148.9, 135.8, 129.9, 123.5, 118.4, 71.1, 61.8, 58.7, 56.3; FAB-MS m/z 280 (M⁺).] Acetyl chloride (0.43 mL, 16.7 mmol) under argon at 0 ⁰C (1 h) was added to an aliquot of this material (494 mg, 1.76 mmol) and anhydrous zinc chloride (9 mg, 0.066 mmol) in anhydrous ether (5.0 mL). Aluminum oxide was added, then the mixture was filtered through a short pad of aluminum oxide and the eluent was evaporated and the residue was purified by silica gel column chromatography to yield 2-(chloromethyl)-5,6-dimethoxybenzenesulfonyl chloride (407 mg, 81.2% yield) as colorless oil [¹H NMR (CDCl₃): δ 123 (d, IH, J = 8.8 Hz), 7.17 (d, IH, J = 8.8 Hz), 4.93 (s, 2H), 4.01 (s, 3H), 3.90 (s, 3H); ¹³C NMR (CDCl₃): δ 154.3, 149.5, 136.3, 128.0, 127.3, 118.2, 61.9, 56.4, 43.6; FAB-MS m/z 283.9 (M⁺).] Triethylamine (86 μL, 0.62 mmol) was added to a solution of this material (88 mg, 0.31 mmol) and hydrazide 19 (64 mg, 0.31 mmol) in anhydrous acetonitrile (2.0 mL) and the resulting mixture was stirred at reflux overnight. The solvent was evaporated and the residue was extracted using dichloromethane. The organic phase was dried (Na₂SO₄) and evaporated and the residue was recrystallized from MeOH to afford 2Oi as a solid (62 mg, 47.8% yield). ¹H NMR (CDCl₃): δ 7.11 (t, 2H, J = 8.4 Hz), 7.02 (d, IH, J = 8.4 Hz), 6.97 (d, IH, / = 8.4 Hz), 4.77 (s, 2H), 4.64 (s, 2H), 4.02 (s, 3H), 3.98 (s, 3H), 3.85 (s, 3H), 3.83 (s, 3H); ¹³C NMR (CDCl₃): δ 165.3, 152.2, 152.1, 147.6, 144.3, 133.1, 127.1, 125.6, 122.2, 119.4, 118.8, 118.1, 117.6, 62.4, 61.8, 56.7 (2C), 50.5, 49.2; FAB-MS m/z 421.1 (MH⁺).

Examples 32-36 General Procedure F for the Synthesis of Hydrazides 21j - 21n. [0123] Triethylamine (1.0 mmol) was added drop-wise to a mixture of appropriately substituted phthalic anhydride (1.0 mmol) and hydrazide 20 (1.0 mmol) in toluene (5.0 mL) and the resulting mixture was stirred at reflux (overnight). The solvent was evaporated and the residue was extracted with dichloromethane. The organic phase was dried (Na₂SO₄) and taken to dryness. Purification of the resulting residue by silica gel column chromatography afforded the desired products 21j - 21n.

Example 32

2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-isoindole-l,3(2H)-dione (21j).

[0124] Following general procedure F using phthalic anhydride as an acylating reagent provided product 21j in 71% yield. ¹H NMR (CDCl₃): δ 7.85-7.83 (m, 2H), 7.76-7.73 (m, 2H), 7.13 (d, IH, J = 8.4 Hz), 7.06 (dd, IH, J = 0.8 Hz, 8.4 Hz), 4.61 (s, 2H), 4.01 (s, 3H), 3.84 (s, 3H); ¹³C NMR (CDCl₃): δ 164.9 (2C), 164.8, 152.3, 147.9, 134.8 (2C), 133.4, 130.0 (2C), 124.0 (2C), 121.4, 118.2, 117.8, 62.5, 56.7, 50.1; FAB-MS m/z 338.9 (MH⁺). Example 33

2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4-hydroxyisoindole-l,3(2H)- dione (21k).

[0125] Following general procedure F using 3-hydroxyphthalic anhydride as an acylating reagent provided product 21k in 83% yield. ¹H NMR (DMSO): δ 11.38 (s, IH), 7.69 (dd, IH, J = 7.6 Hz, 8.4 Hz), 7.41-7.35 (m, 2H), 7.28 (d, 2H, J = 8.4 Hz), 4.57 (s, 2H), 3.82 (s, 6H); ¹³C NMR (DMSO): δ 165.1, 164.8, 163.6, 156.3, 152.3, 147.1, 137.4, 134.0, 131.6, 124.6, 121.3, 119.7, 118.9, 115.2, 113.0, 62.1, 56.9, 50.2; FAB-MS m/z 353.2 (M-H).

Example 34 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5-dimethoxyisoindole- l,3(2H)-dione (211).

[0126] Following general procedure F using 3,4-dimethoxy-phthalic anhydride (Baudart, M. G.; and Hennequin, L. F.. J. Antibiot. 1993, 46, 1458-1470) as an acylating reagent provided product 211 in 63% yield. ¹H NMR (CDCl₃): δ 7.53 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.14-7.11 (m, 2H), 7.05 (dd, IH, J = 0.8 Hz, 8.4 Hz), 4.58 (d, 2H, J = 0.8 Hz), 4.08 (s, 3H), 4.01 (s, 3H), 3.90 (s, 3H), 3.84 (s, 3H); ¹³C NMR (CDCl₃): δ 164.9, 164.2, 163.0, 158.2, 152.2, 147.9, 147.7, 133.4, 122.1, 121.5, 120.3, 119.7, 118.2, 117.7, 116.5, 62.6, 62.5, 56.7, 56.6, 50.1; FAB-MS m/z 399.1 (MH⁺).

Example 35

2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5-dimethoxyisoindole- l,3(2H)-dione (21m).

[0127] Following general procedure F using 4,5-dimethoxy-phthalic anhydride as an acylating reagent provided product 21m in 97% yield. ¹H NMR (CDCl₃): δ 7.31 (s, 2H), 7.14 (d, IH, J = 8.4 Hz), 7.07 (d, IH, J = 8.4 Hz), 4.62 (s, 2H), 4.03 (s, 3H), 3.96 (s, 6H), 3.86 (s, 3H); ¹³C NMR (CDCl₃): δ 165.3 (2C), 164.9, 154.5 (2C), 152.3, 148.0, 133.5, 123.6, 121.5, 118.1 (2C), 117.7 (2C), 105.8 (2C), 62.5, 56.7 (2C), 50.3; FAB-MS m/z 399.1 (MH⁺). Example 36

2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4-fluoroisoindole-l,3(2H)- dione (2In).

[0128] Following general procedure F using 4-fluorophthalic anhydride as an acylating reagent provided product 21n in 96% yield. ¹H NMR (CDCl₃): δ 7.88 (dd, IH, J = 4.4 Hz, 8.0 Hz), 7.54 (dd, IH, J = 2.4 Hz, 6.8 Hz), 7.45-7.40 (m, IH), 7.14 (d, IH, J = 8.4 Hz), 7.07 (d, IH, J = 8.4 Hz), 4.61 (s, 2H), 4.01 (s, 3H), 3.85 (s, 3H); ¹³C NMR (CDCl₃): δ 167.9, 164.7, 163.9, 152.3, 148.0, 133.3, 132.8 (d, 1C, 7= 9.9 Hz), 126.7 (d, 1C, 7= 9.9 Hz), 125.9, 122.1, 121.8, 121.2, 118.2, 117.9, 111.8 (d, 1C, 7 = 25.2 Hz), 62.5, 56.7, 50.1; FAB-MS m/z 357 (MH⁺).

Example 37

2,3-Dihydro-6,7-dimethoxy-2-[(2,3-dihydroxyphenylmethylene)amino]-lH-isoindol-l- one (21o).

[0129] A suspension of hydrazide 20 (248 mg, 1.19 mmol) and 2,3- dihydroxybenzylaldehyde (165 mg, 1.20 mmol) in anhydrous toluene (3 mL) was stirred at reflux (overnight). The product hydrazone 21o was collected by filtration (350 mg, 90% yield). ¹H NMR (DMSO): δ 8.26 (s, IH), 7.27 (dd, 2H, 7 = 8.0 Hz, 25.2 Hz), 6.98 (dd, IH, 7 = 1.6 Hz, 8.0 Hz), 6.81(dd, IH, J = 1.6 Hz, 8.0 Hz), 6.71 (t, IH, 7 = 8.0 Hz), 4.74 (s, 2H), 3.86 (s, 3H), 3.79 (s, 3H); ¹³C NMR (DMSO): δ 162.1, 152.4, 147.1, 146.2, 146.0, 145.4, 132.3, 123.2, 120.6, 119.6, 119.4, 119.3, 118.4, 117.7, 62.1, 56.8, 46.5; FAB-MS m/z 329.1 (M-H). Example 38

2,3-Dihydro-6,7-dimethoxy-2-[(4-fluorophenylmethylene)amino]-lH-Isoindol-l-one (2Ip).

[0130] A suspension of hydrazide 20 (297 mg, 1.43 mmol) and 4-fluorobenzylaldehyde (0.15 mL, 1.44 mmol) in anhydrous toluene (2.5 mL) was stirred at reflux (4 h). The reaction mixture was cooled to room temperature and extracted with dichloromethane and the organic phase was dried (Na₂SO₄) and taken to dryness. Crystallization from EtOAc provided product hydrazone 21p as a white solid (203 mg, 45% yield). ¹H NMR (DMSO): δ 8.13 (s, IH), 7.79 (dd, 2H, J = 5.6 Hz, 8.8 Hz), 7.31-7.22 (m, 4H), 4.72 (s, 2H), 3.84 (s, 3H), 3.79 (s, 3H); ¹³C NMR (DMSO): δ 162.5, 152.4, 147.1, 143.1, 132.3, 131.7, 129.7, 129.6, 123.6, 119.2, 118.2, 116.5, 116.3, 116.1, 62.2, 56.9, 47.2; FAB-MS m/z 315.1 (MH⁺). Examples 42-56

General Procedure G for the Demethylation of Methyl Ethers.

[0131] Boron tribromide (1.0 M in dichloromethane, 8.5 mmol) was added carefully to a solution of appropriate methyl ether (1.0 mmol in 1.0 mL anhydrous dichloromethane) and the mixture was stirred at room temperature (overnight). The reaction was quenced by the addition of ice water (1.0 mL) then the mixture was stirred at room temperature (overnight). The resulting suspension was filtered and the collected solid was purified by preparative HPLC.

[0132] The following were prepared by demethylation of intermediates 21 using General Procedure G. Example 42

/V-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-benzamide (22a).

[0133] Demethylation of 21a according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 25% B to 35% B over 30 minutes; retention time = 17.8 minutes) afford product 22a as a white solid following lyophilization. ¹H NMR

(DMSO): δ 10.83 (s, IH), 7.88 (dd, IH, 7 = 1.6 Hz, 7.6 Hz), 7.58-7.56 (m, IH), 7.51-7.47 (m, 2H), 6.99 (d, IH, J = 8.0 Hz), 6.77 (d, IH, J = 8.0 Hz), 4.42 (s, 2H); ¹³C NMR (DMSO): δ 167.2, 166.1, 145.0, 143.9, 132.7, 132.2, 131.7, 129.0 (2C), 128.0 (2C), 120.3, 116.8, 114.3, 50.8; FAB-MS m/z 283 (M-H). HRMS calcd for C₁₅H₁₃N₂O₄ [MH+]: 285.0875. Found: 285.0871.

Example 43 N-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-(2-hydroxy)benzamide (22b).

[0134] Demethylation of 21b according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 20% B to 50% B over 30 minutes; retention time = 22.5 minutes) afford product 22b as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.60 (s, IH), 10.76 (s, IH), 7.85 (dd, IH, 7 = 1.6 Hz, 8.0 Hz), 7.43 (dt, IH, J =1.6Hz, 8.8 Hz), 6.99 (d, IH, J = 8.0 Hz), 6.96-6.90 (m, 2H), 6.77 (d, IH, J = 8.0 Hz), 4.42 (s, 2H); ¹³C NMR (DMSO): δ 167.9, 167.0, 159.3, 145.1, 143.9, 134.8, 131.8, 129.2, 120.4, 119.6, 117.8, 116.7, 115.1, 114.2, 50.8; FAB-MS m/z 299.1 (M-H). HRMS calcd for C₁₅H₁₃N₂O₅ [MH+]: 301.0824. Found: 301.0834.

Example 44 N-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-(2,3-dihydroxy)benzamide (22c).

[0135] Demethylation of 21c according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 10% B to 40% B over 30 minutes; retention time = 24.1 minutes) afford product 22c as a white solid following lyophilization. ¹H NMR (DMSO): δ 10.91 (bs, IH), 7.30 (d, IH, J = 8.0 Hz), 6.85 (d, IH, / = 8.0 Hz), 6.95 (d, IH, J = 8.0 Hz), 6.77 (d, IH, J = 8.0 Hz), 6.71 (t, IH, J = 8.0 Hz), 4.42 (s, 2H); ¹³C NMR (DMSO): δ 169.1, 167.1, 149.3, 146.7, 145.1, 143.9, 131.8, 120.5, 119.2, 118.4, 118.1, 116.6, 114.5, 114.2, 50.8; FAB-MS m/z 317.2 (MH⁺). HRMS calcd for Ci₅H₁₃N₂O₆ [MH+]: 317.0774. Found: 317.0778.

Example 45 iV-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-4-fluorobenzamide (22d).

[0136] Demethylation of the bis-(4-fluorobenzamide) compound 21d according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 40% B over 30 minutes; retention time = 19.9 minutes) afford the mono-(4-fluorobenzamide) product 22d as a white solid following lyophilization. ¹H NMR (DMSO): δ 10.88 (s, IH), 9.45 (bs, IH), 7.97-7.93 (m, 2H), 7.33 (t, 2H, J = 8.8 Hz), 6.99 (d, IH, J = 8.0 Hz), 6.77 (d, IH, 7 = 8.0 Hz), 4.41 (s, 2H); FAB-MS m/z 303.0 (MH⁺). HRMS calcd for C₁₅H₁₂FN₂O₄ [MH+]: 303.0781. Found: 303.0793.

Example 46 N-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-N-(4-fluorobenzeπesulfonyI)-4- flourobenzesulfonamide (22e)

and iV-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-4-flourobenzesulfonamide

(22f).

[0137] Demethylation of the bis-(4-fluorobenzesulfonamide) compound 21f according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 80% B over 30 minutes) provided the mono-(4-fluorobenzesulfonamide) compound 22e (retention time = 17.8 minutes) as well as the bis-(4-fluorobenzesulfonamide) compound 22f (retention time = 28.5 minutes) as a white solids following lyophilization. Compound 22e: ¹H NMR (DMSO): S 10.59 (s, IH), 9.45 (bs, IH), 8.93 (bs, IH), 7.88-7.85 (m, 2H), 7.37 (t, 2H, J = 8.8 Hz), 6.94 (d, IH, J = 8.0 Hz), 6.70 (d, IH, J = 8.0 Hz), 4.35 (s, 2H); FAB-MS m/z 337 (M-H). HRMS calcd for C₁₄H₁₂FN₂O₅S [MH+]: 339.0451. Found: 339.0454. Compound 22f: ¹H NMR (DMSO): δ 9.66 (bs, IH), 9.38 (bs, IH), 7.98-7.94 (m, 4H), 7.49 (dt, 4H, J = 2.0 Hz, 8.8 Hz), 7.04 (d, IH, 7 = 8.0 Hz), 6.77 (d, IH, J = 8.0 Hz), 4.51 (s, 2H); FAB-MS m/z 495 (M-H). HRMS calcd for C₂₀H₁₅F₂N₂O₇S₂ [MH+]: 497.0289. Found: 497.0299.

Example 47 N-(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-N-(3,4- dihydroxybenzenesuIfonyl)-3,4-dihydroxybenzesuIfonamide (22g).

[0138] Demethylation of compound 21g according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 20% B to 65% B over 30 minutes; retention time = 20.1 minutes) afforded product 22g as a white solid following lyophilization. ¹H ΝMR (DMSO): δ 10.30 (s, 2H), 9.79 (s, 2H), 9.58 (s, IH), 9.30 (s, IH), 7.26 (d, 2H, / = 2.0 Hz), 7.18 (dd, 2H, / = 2.0 Hz, 8.4 Hz), 7.01 (d, IH, J = 8.0 Hz), 6.84 (d, 2H, J = 8.4 Hz), 6.75 (d, IH, /= 8.0 Hz), 4.30 (s, 2H); ¹³C ΝMR (DMSO): δ 166.0, 152.2 (2C), 145.8 (2C), 145.4, 144.5, 131.6, 127.8 (2C), 122.0, 121.1 (2C), 115.9 (2C), 115.8 (2C), 114.6, 114.3, 51.1; FAB-MS m/z 523 (M-H). HRMS calcd for C₂₀Hi₇N₂O₁₁S₂ [MH+]: 525.0274. Found: 525.0285.

Example 48

6,6',7,7^t-Tetrahydroxy-[2,2^t-bi-2H-isoindole]-l,l'(3H,3Η)-dione (22h).

[0139] Demethylation of compound 21h according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 10% B to 35% B over 30 minutes; retention time = 23.2 minutes) afforded product 22g as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.01 (d, 2H, J = 8.0 Hz), 6.79 (d, 2H, J = 8.0 Hz), 4.49 (s, 4H); ¹³C NMR (DMSO): δ 166.6 (2C), 145.2 (2C), 144.0 (2C), 131.8 (2C), 120.5 (2C), 116.3 (2C), 114.3 (2C), 49.3 (2C); FAB-MS m/z 328.28 (MH⁺). HRMS calcd for C₁₆H₁₃N₂O₆ [MH+]: 329.0774. Found: 329.0776.

Example 49

N-tCl^-Dihydro-όjV-dihydroxy-l-oxo^H-isoindol-Z-yD-iV'-tCδ^-dihydroxy-l.l-dioxido- l,2-benzisothiazol-2(3H)-yl)] Hydrazide (22i).

[0140] Demethylation of 21i according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 10% B to 35% B over 30 minutes; retention time = 27.8 minutes) afforded product 22i as a white solid following lyophilization. ¹H NMR (DMSO): δ 10.18 (s, IH), 10.10 (s, IH), 9.47 (s, IH), 9.08 (s, IH), 7.07 (d, IH, J = 8.0 Hz), 6.99 (d, IH, J = 8.0 Hz), 6.77-6.73 (m, 2H), 4.63 (s, 2H), 4.52 (s, 2H); ¹³C NMR (DMSO): δ 166.8, 145.6, 145.1, 144.0, 141.9, 131.6, 124.5, 121.3, 120.7, 115.9, 115.4, 114.3, 48.9; FAB- MS m/z 363 (M-H). HRMS calcd for C₁₅H₁₃N₂O₇S [MH+]: 365.0443. Found: 365.0444.

Example 50 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-isoindole-l,3(2H)-dione (22j).

[0141] Demethylation of 21j according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 45% B over 30 minutes; retention time = 22.2 minutes) afforded product 22j as a white solid following lyophilization. ¹H NMR (DMSO): δ 9.64 (s, IH), 9.30 (s, IH), 7.98-7.95 (m, 2H), 7.94-7.91 (m, 2H), 7.06 (d, IH, J = 8.0 Hz), 6.83 (d, IH, / = 8.0 Hz), 4.53 (s, 2H); ¹³C NMR (DMSO): δ 166.2, 165.3 (2C), 145.5, 144.5, 135.9 (2C), 132.2, 129.9 (2C), 124.3 (2C), 120.9, 115.3, 114.5, 50.3; FAB-MS m/z 309 (M-H). HRMS calcd for Ci₆Hi₁N₂O₅ [MH+]: 311.0668. Found: 311.0674.

Example 51 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-3-hydroxyisoindole-l,3(2H)- dione (22k).

[0142] Demethylation of 21k according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 100% B over 30 minutes; retention time = 19.3 minutes) afforded product 22k as a white solid following lyophilization. ¹H NMR (DMSO): <H 1.35 (s, IH), 9.61 (s, IH), 9.26 (s, IH), 7.68 (dt, IH, 7 = 1.6 Hz, 8.4 Hz), 7.36 (d, IH, J = 8.4 Hz), 7.28 (d, IH, J = 8.4 Hz), 7.05 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 6.81 (d, IH, 7 = 8.0 Hz), 4.49 (s, 2H); ¹³C NMR (DMSO): δ 166.2, 165.2, 163.8, 156.2, 145.5, 144.4, 137.4, 132.1, 131.6, 124.5, 120.8, 115.5, 115.2, 114.4, 113.1, 50.3; FAB-MS m/z 325.1 (M- H). HRMS calcd for Ci₆H₁₁N₂O₆ [MH+]: 327.0617. Found: 327.0621. Example 52

2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-3,4-dihydroxyisoindole- l,3(2H)-dione (221).

[0143] Demethylation of 211 according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 10% B to 35% B over 30 minutes; retention time = 23.5 minutes) afforded product 221 as a white solid following lyophilization. ¹H NMR (DMSO): δ 11.11 (s, IH), 10.44 (s, IH), 9.59 (s, IH), 9.23 (s, IH), 7.26 (d, IH, 7 = 8.0 Hz), 7.11 (d, IH, J = 8.0 Hz), 7.04 (d, IH, 7 = 8.0 Hz), 6.81 (d, IH, J = 8.0 Hz), 4.47 (s, 2H); ¹³C NMR (DMSO): δ 166.2, 165.0, 163.9, 153.5, 145.4 (2C), 144.3, 132.1, 120.7, 120.3, 119.7, 117.1, 115.6, 114.4, 114.1, 50.3. FAB-MS m/z 340.9 (M-H). HRMS calcd for Ci₆H_nN₂O₇ [MH+]: 343.0566. Found: 343.0566.

Example 53

2-[l,3-Dihydro-6,7-dihydoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5-dihydroxyisoindole- l,3(2H)-dione (22m).

[0144] Demethylation of 21m according to general procedure G and purification by preparative HPLC [Phe] (linear gradient of 20% B to 30% B over 30 minutes; retention time = 22.5 minutes) afforded product 22m as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.19 (s, 2H), 7.05 (d, IH, 7 = 8.0 Hz), 6.80 (d, IH, J = 8.0 Hz), 4.48 (s, 2H); ¹³C NMR (DMSO): δ 166.2, 165.5 (2C), 152.2 (2C), 145.4, 144.4, 132.1, 122.0 (2C), 120.8, 115.5, 114.4, 110.7 (2C), 50.5; FAB-MS m/z 341 (M-H). HRMS calcd for Ci₆H_nN₂O₇ [MH+]: 343.0566. Found: 343.0560.

Example 54 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-4-fluoroisoindole-l,3(2H)- dione (22n).

[0145] Demethylation of 21n according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 50% B over 30 minutes; retention time = 25.3 minutes) afforded product 22n as a white solid following lyophilization. ¹H NMR (DMSO): δ 9.64 (s, IH), 9.31 (s, IH), 8.06 (dd, IH, J = 4.8 Hz, 8.4 Hz), 7.91 (dd, IH, J = 4.8 Hz, 7.6 Hz), 7.78-7.73 (m, IH), 7.06 (d, IH, 7 = 8.0 Hz), 6.83 (d, IH, 7 = 8.0 Hz), 4.52 (s, 2H); ¹³C NMR (DMSO): δ 166.2, 165.3, 164.4, 145.5, 144.5, 132.9 (d, 1C, 7 = 9.9 Hz), 132.2, 127.4 (d, 1C, 7 = 9.9 Hz), 126.2, 122.8, 122.6, 120.9, 115.2, 114.5, 112.4 (d, 1C, 7 = 25.1 Hz), 50.2; FAB-MS m/z 327 ^'.1 (M-H). HRMS calcd for C₁₆H₁₀FN₂O₅ [MH+]: 329.0574. Found: 329.0581.

Example 55

2,3-Dihydro-6,7-dihydroxy-2-[(2,3-dihydroxyphenylmethylene)amino]-lH-isoindol-l- one (22o).

[0146] Demethylation of 21o according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 20% B to 50% B over 30 minutes; retention time = 22.3 minutes) afforded product 22o as a white solid following lyophilization. ¹H NMR (DMSO): δ 8.21 (s, IH), 7.00 (d, IH, J = 7.6 Hz), 6.96 (dd, IH, J= 1.6 Hz, 8.0 Hz), 6.82- 6.79 (m, 2H), 6.71 (t, IH, 7 = 8.0 Hz), 4.70 (s, 2H); ¹³C NMR (DMSO): δ 163.9, 146.1, 146.0, 145.4, 144.9 (2C), 144.4, 130.2, 120.7, 119.7, 119.5, 117.6, 117.0, 114.3, 46.6; FAB- MS m/z 299 (M-H). HRMS calcd for C₁₅H₁₃N₂O₅ [MH+]: 301.0824. Found: 301.0829.

Example 56 2,3-Dihydro-6,7-dihydroxy-2-[(4-fluorophenylmethylene)amino]-lH-isoindol-l-one (22p).

[0147] Demethylation of 21p according to general procedure G and purification by preparative HPLC [YMC] (linear gradient of 30% B to 55% B over 30 minutes; retention time = 20.0 minutes) afforded product 22p as a white solid following lyophilization. ¹H NMR (DMSO): δ 8.09 (s, IH), 7.78 (dd, 2H, 7 = 1.6 Hz, 8.4 Hz), 7.26 (t, 2H, 7 = 8.4 Hz), 6.99 (d, IH, 7 = 8.0 Hz), 6.80 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 4.67 (s, 2H); ¹³C NMR (DMSO): δ 164.6, 164.4, 162.1, 145.3, 144.3, 142.4, 131.8 (d, 1C, 7 = 3.0 Hz), 130.0, 129.6 (d, 1C, 7 = 8.4 Hz), 120.7, 117.4, 116.5 (d, 1C, 7 = 8.4 Hz), 116.2 (d, 1C, 7 = 8.4 Hz), 114.2, 47.4; FAB-MS m/z 287.1 (MH⁺). HRMS calcd for C₁₅H₁₂FN₂O₃ [MH+]: 287.0832. Found: 287.0832.

Examples 57-61 General Procedure H for the Synthesis of Amides 23a - e.

[0148] Triethylamine (2.0 mmol) was added to a solution of methyl 2-chloromethyl-3,4- dimethoxybenzoate (19) (1.0 mmol) and appropriate amine (1.0 mmol) in anhydrous acetonitrile (3.0 mL) was added. The mixture was stirred at reflux until the starting material was consumed as indicated by silica gel TLC. The solvent was evaporated and the residue was partitioned between chloroform and brine. The combined organic phase was dried (Na₂SO₄), evaporated and the residue was purified by silica gel column chromatography.

Example 57 2,3-Dihydro-6,7-dimethoxy-2-phenylmethyl-H-isoindol-l-one (23a).

[0149] Reaction of benzylamine according to general procedure H provided 23a in 67% yield. ¹H NMR (CDCl₃): δ 7.21-7.18 (m, 4H), 7.17-7.15 (m, IH), 6. 96 (d, IH, J = 8.0 Hz), 6.89 (d, IH, J = 8.0 Hz), 4.63 (s, 2H), 4.04 (s, 2H), 4.02 (d, 3H, J = 1.2 Hz), 3.77 (d, 3H, J = 1.2 Hz); ¹³C NMR (CDCl₃): δ 166.6, 152.2, 147.2, 137.1, 134.5, 128.6 (2C), 128.1 (2C), 127.5, 124.7, 117.8, 116.4, 62.4, 56.7, 48.4, 46.2; FAB-MS m/z 284.2 (MH⁺).

Example 58

2,3-Dihydro-6,7-dimethoxy-2-(2-phenylethyl)-H-isoindol-l-one (23b).

[0150] Reaction of 2-phenylethylamine according to general procedure H provided 23b in 87% yield. ¹H NMR (CDCl₃): δ 7.18-7.10 (m, 5H), 6.96 (d, IH, 7 = 8.4 Hz), 6.91 (d, IH, J = 8.4 Hz), 4.01 (s, 2H), 3.99 (s, 3H), 3.77 (s, 3H), 3.70 (t, 2H, J = 7.2 Hz), 2.87 (t, 2H, J = 7.2 Hz); ¹³C NMR (CDCl₃): δ 166.6, 152.2, 147.0, 138.8, 134.5, 128.6 (2C), 128.5 (2C), 126.4, 125.0, 117.7, 116.3, 62.4, 56.7, 49.5, 44.1, 34.6; FAB-MS m/z 298.1 (MH⁺).

Example 59 2,3-Dihydro-6,7-dimethoxy-2-(l-naphthylmethyl)-H-isoindol-l-one (23c).

[0151] Reaction of 1-naphthylmethylamine according to general procedure H provided 23c in 58% yield. ¹H NMR (CDCl₃): δ 8.19 (d, IH, J = 8.4 Hz), 7.76-7.72 (m, 2H), 7.47-7.43 (m, IH), 7.40-7.33 (m, 3H), 6.88 (dd, IH, J = 6.8 Hz, 8.4 Hz), 6.76 (dd, IH, J = 6.8 Hz, 8.4 Hz), 5.08 (d, 2H, / = 1.6 Hz), 4.08 (d, 3H, J = 1.2 Hz), 3.88 (d, 2H, J = 2.0 Hz), 3.75 (d, 3H, J = 2.4 Hz); ¹³C NMR (CDCl₃): δ 166.2, 152.2, 147.2, 134.4, 133.8, 132.6, 131.5, 128.8, 128.5, 127.5, 126.8, 126.0, 125.2, 124.7, 123.9, 117.8, 116.4, 64.5, 56.6, 48.4, 44.5; FAB-MS m/z 334.1 (MH⁺).

Example 60 2,3-Dihydro-6,7-dimethoxy-2-(4-fluorophenylmethyl)-H-isoindoI-l-one (23d).

[0152] Reaction of 4-fluorophenylmethylamine according to general procedure H provided 23d in 69% yield. ¹H NMR (CDCl₃): δ 1.26-1.23 (m, 2H), 7.03 (d, IH, J = 8.0 Hz), 6.98-6.94 (m, 3H), 4.67 (s, 2H), 4.11 (s, 2H), 4.07 (s, 3H), 3.84 (s, 3H); ¹³C NMR (CDCl₃): δ 166.6, 163.4, 161.0, 152.4, 147.4, 134.4, 132.9 (d, 1C, 7 = 3.0 Hz), 129.9 (d, 1C, J = 8.4 Hz), 124.7, 117.7, 116.5, 115.7, 115.4, 62.5, 56.8, 48.4, 45.6; FAB-MS m/z 302.1 (MH⁺).

Example 61 2,3-Dihydro-6,7-dimethoxy-2-[(3-chloro-4-fluorophenyl)methyl]-H-isoindol-l-one (23e).

[0153] Reaction of [(3-chloro-4-fluorophenyl)methyl]amine according to general procedure H provided 23e in 21% yield. ¹H NMR (CDCl₃): δ 7.29 (dd, IH, J = 2.0 Hz, 7.6 Hz), 7.14- 7.11 (m, IH), 7.02 (d, IH, J = 8.4 Hz), 7.01 (s, IH), 4.61 (s, 2H), 4.12 (s, 2H), 4.04 (s, 3H), 3.82 (s, 3H); ¹³C NMR (CDCl₃): δ 166.1, 158.7, 156.2, 152.4, 147.3, 134.3 (d, 1C, J = 3.8 Hz), 130.2, 127.9 (d, 1C, J = 7.6 Hz), 124.4, 121.1 (d, 1C, J = 17.6 Hz), 117.9, 116.9, 116.7, 62.4, 56.7, 48.5, 45.2; FAB-MS m/z 336.1 (MH⁺).

Example 62 2,3-Dihydro-6,7-dimethoxy-2-[3-((hydroxymethyl)phenyl)methyl]-H-isoindol-l-one

(23f).

[0154] Lithium aluminum hydride (1.0 M in THF, 29.9 mL, 29.9 mmol) was added slowly to a solution of 3-cyanobenzaldehyde (1.12 g, 9.25 mmol) in anhydrous THF (25.0 mL) at room temperature under argon and the mixture was stirred at reflux (overnight). The reaction mixture was cooled to room temperature and quenched by addition of ice and NaOH (aq. 3.0 N, 10.0 mL). The mixture was extracted with chloroform (3 x 80 mL) and the combined organic phase was washed with brine (2 x 20 mL) and dried (Na₂SO₄). The solvent was evaporated and the residue was purified by silica gel column chromatography to yield [3- (hydroxymethyl)phenyl] amine as colorless oil (70% yield). [¹H NMR (CDCl₃): δ 7.26-7.22 (m, IH), 7.21-7.19 (m, IH), 7.16-7.14 (m, IH), 7.12-7.09 (m, IH), 4.55 (d, 2H, J = 6.0 Hz), 3.71 (d, 2H, /= 8.0 Hz); ¹³C NMR (CDCl₃): δ 142.7, 142.0, 128.6, 126.0, 125.5, 125.4, 64.4, 46.1; FAB-MS m/z 138.1 (MH⁺).] Reaction of this material according to general procedure G provided 23f in 38% yield. ¹H NMR (CDCl₃): δ 7.22 (s, IH), 7.20-7.18 (m, 2H), 7.11-7.09 (m, IH), 6. 98 (d, IH, 7 = 8.4 Hz), 6.91 (d, IH, 7 = 8.4 Hz), 4.60 (s, 2H), 4.57 (s, 2H), 4.06 (s, 2H), 4.01 (d, 3H, 7 = 1.2 Hz), 3.80 (d, 3H, 7 = 1.2 Hz); ¹³C NMR (CDCl₃): δ 166.8, 152.2, 147.2, 141.9, 137.1, 134.5, 128.8, 127.0, 126.6, 126.1, 124.7, 117.8, 116.5, 64.6, 62.4, 56.7, 48.5, 46.2; FAB-MS m/z 314.1 (MH⁺). Example 63

2,3-Dihydro-6,7-dimethoxy-2-[3-((phenylmethyl)phenyl)methyl]-H-isoindol-l-one (23g).

[0155] Tetrakis(triphenylphosphine)palladium(0) (354 mg, 0.306 mmol) was added under argon to a mixture of (3-benzyl)phenyl bromide (688 mg, 2.49 mmol) and zinc cyanide (1.96 g, 16.7 mmol) (Wai, J. S. et al. J. Med. Chem. 2000, 43, 4923-4926) in dimethylformamide (5.0 mL) and the resulting mixture was stirred under argon at 95 ^°C (2 d). The mixture was diluted with ethyl acetate and washed successively with H₂O, dilute aqueous HCl acid and brine. The organic phase was dried (Na₂SO₄) and the solvent removed under vacuum. The residue was purified by silica column chromatography (ethyl acetate-hexanes) to yield 2- cyano-4-methylbiphenyl as a colorless oil (369 mg, 69% yield). [¹H NMR (CDCl₃): δ 7.48- 7.45 (m, 2H), 7.43-7 '.41 (m, IH), 7.38-7.30 (m, 3H), 7.26-7.22 (m, IH), 7.17-7.12 (m, 2H), 3.99 (s, 2H); ¹³C NMR (CDCl₃): δ 142.6, 139.5, 133.4, 132.3, 129.9, 129.3, 128.9 (2C),

128.8 (2C), 126.7, 118.9, 112.5, 41.4; FAB-MS m/z 193 (M⁺).] Lithium aluminum hydride (1.0 M in THF, 5.35 mL, 5.35 mmol) was added to a solution of this material (344 mg, 1.78 mmol) in anhydrous THF (5.0 mL) at room temperature under argon and the mixture was stirred at reflux (4 h). The mixture was cooled to room temperature and quenched by addition of aqueous NaOH (3.0 N, 10.0 mL). The mixture was extracted with ethyl acetate (3 x 80 mL) and the combined organic phase was washed with brine (2 x 20 mL) and dried (Na₂SO₄). The solvent was evaporated and the residue was purified by silica gel column chromatography to provide [3-((phenylmethyl)phenyl)methyl]amine as a colorless oil (94 mg, 27% yield) [¹H NMR (CDCl₃): δ 7.31-7.25 (m, 3H), 7.24-7.19 (m, 3H), 7.15-7.14 (m, 2H), 7.09-7.07 (m, IH), 3.98 (s, 2H), 3.81 (s, 2H); ¹³C NMR (CDCl₃): δ 143.2, 141.4, 141.1,

128.9 (2C), 128.7, 128.5 (2C), 127.7, 127.5, 126.1, 124.9, 46.3, 41.9; FAB-MS m/z 198.1 (MH⁺).] Treatment of this material according to general procedure G provided 23g in 76% yield. ¹H NMR (CDCl₃): δ 7.25-7.20 (m, 3H), 7.18-7.16 (m, IH), 7.15-7.13 (m, 2H), 7.12- 7.09 (m, 2H) 7.06-7.04 (m, IH), 7.03 (d, IH, J = 8.4 Hz), 6.96 (d, IH, J = 8.4 Hz), 4.68 (s, 2H), 4.10 (s, 2H), 4.09 (s, 3H), 3.92 (s, 2H), 3.86 (s, 3H); ¹³C NMR (CDCl₃): δ 166.6, 152.3, 147.4, 141.7, 140.8, 137.2, 134.5, 128.8 (3C), 128.4 (2C), 128.2, 126.1, 126.0, 124.9, 117.7, 116.5, 62.6, 56.8, 48.5, 46.3, 41.7.

Examples 64-108

[0156] The following were prepared by demethylation of intermediates 23 using General Procedure G.

Example 64 2,3-Dihydro-6,7-dihydroxy-2-phenylmethyl-H-isoindol-l-one (24a).

[0157] Treatment of 23a with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 30% B to 55% B over 30 minutes; retention time = 22.5 minutes) afford product 24a as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.30-7.22 (m, 5H), 6.92 (d, IH, J = 7.6 Hz), 6.69 (d, IH, / = 7.6 Hz), 4.65 (s, 2H), 4.12 (s, 2H); ¹³C NMR (DMSO): δ 169.6, 144.1, 143.0, 136.9, 132.5, 128.4 (2C), 127.5 (2C), 127.3, 119.6, 117.4, 113.6, 49.0, 45.4; FAB-MS m/z 254.1 (M-H). HRMS calcd for Ci₅Hi₄NO₃ [MH+]: 256.0974. Found: 256.0974.

Example 65 2,3-Dihydro-6,7-dihydroxy-2-(2-phenylethyl)-H-isoindol-l-one (24b).

[0158] Treatment of 23b with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 50% B over 30 minutes; retention time = 16.8 minutes) afford product 24b as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.23-7.14 (m, 5H), 6.89 (d, IH, J = 8.0 Hz), 6.70 (d, IH, J = 8.0 Hz), 4.17 (s, 2H), 3.64 (t, 2H, J = 7.2 Hz), 2.84 (t, 2H, J = 7.2 Hz); ¹³C NMR (DMSO): δ 168.4, 144.6, 143.2, 139.4, 132.5, 129.0 (2C), 128.8 (2C), 126.7, 119.9, 118.5, 114.0, 49.6, 43.2, 34.3; FAB-MS m/z 270.1 (MH⁺). HRMS calcd for Ci₆H,₆NO₃ [MH+]: 270.1130. Found: 270.1133. Example 66

2,3-Dihydro-6,7-dihydroxy-2-(l-naphthylmethyl)-H-isoindol-l-one (24c).

[0159] Treatment of 23c with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 60% B over 30 minutes; retention time = 23.9 minutes) afford product 24c as a white solid following lyophilization.Η . ¹H NMR (CD₃OD): δ 8.15 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.86-7.80 (m, 2H), 7.50-7.40 (m, 4H), 6. 89 (d, IH, J = 8.0 Hz), 6.63 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 5.13 (s, 2H), 4.01 (s, 2H); ¹³C NMR (DMSO): δ 167.9, 144.8, 143.4, 133.9, 133.2, 132.6, 131.3, 129.0, 128.7, 127.3, 127.0, 126.4, 125.9, 123.8, 119.9, 118.3, 114.2, 49.0, 43.6; FAB-MS m/z 306.1(MH⁺). HRMS calcd for Ci₉H]₆NO₃ [MH+]: 306.1130. Found: 306.1128.

Example 67

2,3-Dihydro-6,7-dihydroxy-2-(4-fluorophenylmethyI)-H-isoindol-l-one (24d).

[0160] Treatment of 23d with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 30% B to 50% B over 30 minutes; retention time = 24.3 minutes) afford product 24d as a white solid following lyophilization. ¹H NMR (CD₃OD): δ 7.26 (dd, 2H, J = 5.6 Hz, 8.8 Hz), 7.01 (t, 2H, / = 8.8 Hz), 6.92 (d, IH, J = 8.0 Hz), 6.70 (d, IH, J = 8.0 Hz), 4.64 (s, 2H), 4.14 (s, 2H); ¹³C NMR (CDCl₃): δ 169.7, 163.6, 161.1, 143.3, 142.1, 132.3 (d, 1C, J = 3.0 Hz), 131.9, 129.8 (d, 1C, 7 = 8.4 Hz), 119.8, 117.3, 115.8, 115.6, 114.4, 49.6, 45.4; FAB-MS m/z 274.1 (MH⁺). HRMS calcd for Ci₅H]₃FNO₃ [MH+]: 274.0879. Found: 274.0884.

Example 68 2,3-Dihydro-6,7-dihydroxy-2-[(3-chIoro-4-fluorophenyl)methyl]-H-isoindol-l-one (24e).

[0161] Treatment of 23e with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 70% B over 30 minutes; retention time = 18.2 minutes) afford product 24e as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.43 (dd, IH, J = 2.0 Hz, 7.6 Hz), 7.33 (t, IH, 7 = 8.8 Hz), 7.15-7.21 (m, IH), 6.91 (d, IH, 7 = 7.6 Hz), 6.71 (d, IH, 7 = 7.6 Hz), 4.58 (s, 2H), 4.17 (s, 2H); ¹³C NMR (DMSO): δ 168.4, 158.1, 155.7, 144.9, 143.4, 136.0 (d, 1C, J = 3.8 Hz), 132.7, 130.2, 128.8 (d, 1C, J = 7.6 Hz), 120.0, 118.2, 117.5 (d, 1C, J = 21.3 Hz), 114.2, 49.1, 44.5; FAB-MS m/z 308.0 (MH⁺). HRMS calcd for C₁₅H₁₂ClFNO₃ [MH+]: 308.0490. Found: 308.0489.

Example 69 2,3-Dihydro-6,7-dihydroxy-2-[3-((bromomethyl)phenyl)methyI]-H-isoindol-l-one (24f).

[0162] Treatment of 23f with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 40% B to 50% B over 30 minutes; retention time = 23.4 minutes) afford product 24f as a white solid following lyophilization. ¹H NMR (DMSO): <5 7.30-7.29 (m, 3H), 7.17-7.15 (m, IH), 6.91 (d, IH, 7 = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.63 (s, 2H), 4.59 (s, 2H), 4.15 (s, 2H); ¹³C NMR (DMSO): δ 168.4, 144.8, 143.4, 138.9, 138.5, 132.6, 129.5, 128.7 (2C), 128.0, 120.0, 118.2, 114.2, 49.1, 45.3, 34.7; FAB-MS m/z 346 (M-H). HRMS calcd for C₁₆Hi₅BrNO₃ [MH+]: 348.0235. Found: 348.0227.

Example 70 2,3-Dihydro-6,7-dihydroxy-2-[3-((phenylmethyl)phenyl)methyl]-H-isoindol-l-one (24g).

[0163] Treatment of 23g with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 50% B to 65% B over 30 minutes; retention time = 20.5 minutes) afford product 24g as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.23-7.19 (m, 3H), 7.16-7.13 (m, 2H), 7.11-7.06 (m, 3H), 7.03-7.01 (m, IH), 6.90 (d, IH, J = 8.0 Hz), 6.70 (d, IH, 7 = 8.0 Hz), 4.55 (s, 2H), 4.11 (s, 2H), 3.87 (s, 2H); ¹³C NMR (DMSO): S 168.4, 144.8, 143.4, 142.1, 141.5, 138.1, 132.6, 131.7, 131.3, 129.2, 129.1, 128.8, 128.4, 128.2, 126.4, 125.7, 119.9, 118.3, 114.2, 49.1, 45.5, 41.4; FAB-MS m/z 346.1 (MH⁺). HRMS calcd for C₂₂H₂₀NO₃ [MH+]: 346.1443. Found: 346.1450.

Example 71 4,5-Dihydroxy-2-[(3-fluorophenyl)methyl]-H-isoindoI-l-one (24h).

[0164] Linear gradient of 30% B to 60% B over 30 minutes; retention time = 22.3 minutes. ¹H NMR (DMSO): δ 7.36-7.30 (m, IH), 7.07-7.02 (m, 3H), 6.92 (d, IH, J= 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.61 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.5, 162.7 (d, 1C, 7 = 242.6 Hz), 144.8, 143.4, 141.0 (d, 1C, J = 6.9 Hz), 132.7, 131.1 (d, 1C, 7 = 8.4 Hz), 124.0 (d, 1C, 7 = 2.2 Hz), 120.0, 118.2, 114.8 (d, 1C, 7 = 21.3 Hz), 114.5 (d, 1C, 7 = 21.3 Hz), 114.2, 49.2, 45.1. APCI-MS m/z: 274.1 (MH⁺). HRMS calcd for C]₅Hi₃NO₃F [MH+]: 274.0879. Found: 274.0881.

Example 72 4,5-Dihydroxy-2-[(2-fluorophenyl)methyl]-H-isoindol-l-one (24i).

[0165] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 17.9 minutes. ¹H NMR (DMSO): δ 7.31-7.27 (m, IH), 7.26-7.21 (m, IH), 7.17-7.09 (m, 2H), 6.92 (d, IH, 7 = 8.0 Hz), 6.71 (d, IH, 7 = 8.0 Hz), 4.65 (s, 2H), 4.16 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 160.7 (d, 1C, 7 = 244.4 Hz), 144.8, 143.4, 132.6, 130.6 (d, 1C, 7 = 3.8 Hz), 130.0 (d, 1C, 7 = 7.6 Hz), 125.1 (d, 1C, 7 = 3.1 Hz), 124.5 (d, 1C, 7 = 15.5 Hz), 120.0, 118.1, 115.8 (d, 1C, J = 21.4 Hz), 114.2, 49.2, 39.5. APCI-MS m/z: 274.1 (MH⁺). HRMS calcd for C₁₅H]₃NO₃F [MH+]: 274.0879. Found: 274.0883.

Example 73 4,5-Dihydroxy-2-[(3-(azidomethyl)phenyI)methyl]-H-isoindoI-l-one (24j)

[0166] Sodium azide (20 mg, 0.31 mmol) was added to the solution of 2,3-Dihydro-6,7- dihydroxy-2-[3-((bromomethyl)phenyl)methyl]-H-isoindol-l-one¹ (52 mg, 0.15 mmol) in acetone-water (5/1, 1.2 mL). The mixture was refluxed overnight and purified by prepared HPLC. Linear gradient of 40% B to 50% B over 30 minutes; retention time = 20.2 minutes.White solide (15 mg, 31% yield) was afforded. ¹H NMR (CD₃OD): δ 1.31-133 (m, IH), 7.26 (brs, 2H), 7.24 (brs, IH), 6.94 (d, IH, / = 8.0 Hz), 6.73 (dt, IH, / = 0.8 Hz, 8.0 Hz), 4.71 (s, 2H), 4.32 (s, 2H), 4.19 (s, 2H). ¹³C NMR (CD₃OD): δ 169.6, 144.2, 143.1, 137.7, 136.4, 132.5, 128.9, 127.4 (2C), 127.3, 119.6, 117.3, 113.6, 53.9, 49.0, 45.2. APCI-MS m/v 311.1 (MH⁺).

Example 74 4,5-Dihydroxy-2-[(3-chlorophenyl)methyl]-H-isoindol-l-one (24k).

Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.3 minutes. ¹H NMR (DMSO): δ 7.32 (t, IH, J = 7.6 Hz), 7.28 (d, 2H, J = 7.6 Hz), 7.17 (d, IH, / = 7.6 Hz), 6.92 (dd, IH, /= 0.8 Hz, 8.0 Hz), 6.71 (dd, IH, J = 0.8 Hz, 8.0 Hz), 4.60 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 144.9, 143.4, 140.6, 133.7, 132.7, 131.0, 127.9, 127.7, 126.7, 120.0, 118.2, 114.2, 49.2, 45.0. APCI-MS m/z: 290.0 (M⁺). HRMS calcd for C]₅H₁₃NO₃Cl [MH⁺]: 290.0584. Found: 290.0580. Example 75

4,5-Dihydroxy-2-[(3-bromophenyl)methyl]-H-isoindol-l-one (241).

[0167] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 23.9 minutes. ¹H NMR (DMSO): δ 7.42-7.41 (m, 2H), 7.27-7.20 (m, 2H), 6.92 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J= 8.0 Hz), 4.59 (s, 2H), 4.16 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 144.9, 143.4, 140.9, 132.7, 131.3, 130.8, 130.6, 127.1, 122.3, 120.0, 118.2, 114.2, 49.2, 45.0. APCI-MS m/z: 334.0 (M⁺). HRMS calcd for C₅H₁₃NO₃Br [MH⁺]: 334.0079. Found: 334.0074.

Example 76 4,5-Dihydroxy-2-[(3-iodophenyl)methyl]-H-isoindol-l-one (24m).

[0168] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 23.6 minutes. ¹H NMR (DMSO): δ 7.59 (d, IH, J = 1.2 Hz), 7.58 (d, IH, J = 1.2 Hz), 7.22 (dd, IH, J = 0.8 Hz, 7.2 Hz), 7.09 (t, IH, / = 7.2 Hz), 6.92 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.56 (s, 2H), 4.15 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 144.9, 143.4, 140.7, 136.6, 136.5, 132.6, 131.3, 127.5, 120.0, 118.2, 114.2, 95.5, 49.2, 44.9. APCI-MS m/z: 381.9 (MH⁺). HRMS calcd for C₁₅H₁₃NO₃I [MH⁺]: 381.9940. Found: 381.9942.

Example 77 4,5-Dihydroxy-2-[(3, 4-difluorophenyl)methyl]-H-isoindol-l-one (24n).

[0169] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 20.0 minutes. ¹H NMR (DMSO): δ 7.38-7.25 (m, 2H), 7.08-7.05 (m, IH), 6.91 (dd, IH, J = 0.8 Hz, 8.0 Hz), 6.71 (d, IH, J = 0.8 Hz, 8.0 Hz), 4.58 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 154.8 (dd, 1C, J = 3.0 Hz, 244.9 Hz), 154.1 (dd, 1C, 7 = 3.0 Hz, 244.1 Hz), 144.8, 143.4, 135.8 (dd, 1C, 7 = 3.0 Hz, 5.4 Hz), 132.7, 124.9 (dd, 1C, 7 = 3.1 Hz, 6.9 Hz), 120.0, 118.2, 118.1 (d, 1C, 7 = 17.6 Hz), 117.2 (d, 1C, 7 = 17.6 Hz), 114.2, 49.1, 44.6. APCI-MS m/z: 292.1 (MH⁺). HRMS calcd for C₁₅H₁₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0786. Example 78

4,5-Dihydroxy-2-[(3,5-difluorophenyl)methyl]-H-isoindol-l-one (24o).

[0170] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 21.0 minutes. ¹H NMR (DMSO): δ 7.09-7.03 (m, IH), 6.93 (d, IH, 7 = 8.0 Hz), 6.92-6.89 (m, 2H), 6.72 (d, IH, 7 = 8.0 Hz), 4.61 (s, 2H), 4.20 (s, 2H). ¹³C NMR (DMSO): δ 168.5, 163.0 (d, 1C, 7 = 245.6 Hz), 162.9 (d, 1C, 7 = 244.9 Hz), 144.9, 143.5, 142.8 (t, 1C, 7 = 3.8 Hz), 132.8, 120.0, 118.1, 114.2, 111.0 (dd, 2C, 7 = 6.9 Hz, 18.3 Hz), 103.1 (t, 1C, 7 = 25.6 Hz), 49.3, 45.0. APCI-MS m/z: 292.1 (MH⁺). HRMS calcd for Ci₅H₁₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0781. Example 79

4,5-Dihydroxy-2-[(2,5-difluorophenyl)methyl]-H-isoindol-l-one (24p).

[0171] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 19.3 minutes.

¹H NMR (DMSO): δ 7.25-7.20 (m, IH), 7.17-7.11 (m, IH), 7.08-7.04 (m, IH), 6.91 (d, IH, 7 = 8.0 Hz), 6.72 (d, IH, 7 = 8.0 Hz), 4.63 (s, 2H), 4.21 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 158.6 (dd, 1C, 7 = 2.3 Hz, 239.5 Hz), 156.8 (d, 1C, 7 = 238.0 Hz), 144.9, 143.4, 132.8, 126.7 (dd, 1C, 7 = 7.6 Hz, 17.5 Hz), 120.0, 118.0, 117.5 (dd, 1C, 7 = 8.4 Hz, 24.4 Hz), 116.8 (dd, 1C, 7 = 5.4 Hz, 24.4 Hz), 116.3 (dd, 1C, 7 = 8.4 Hz, 24.4 Hz), 114.2, 49.3, 39.5. APCI-MS m/z: 292.1 (MH⁺). HRMS calcd for Ci₅H₁₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0784. Example 80 4,5-Dihydroxy-2-[(2,6-difluorophenyI)methyl]-H-isoindol-l-one (24q).

[0172] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 17.7 minutes. ¹H NMR (DMSO): δ 7.42-7.34 (m, IH), 7.09-7.05 (m, 2H), 6.89 (d, IH, / = 8.0 Hz), 6.70 (d, IH, J= 8.0 Hz), 4.68 (s, 2H), 4.13 (s, 2H). ¹³C NMR (DMSO): δ 167.8, 161.5 (d, 1C, 7 = 246.4 Hz), 161.4 (d, 1C, J = 246.4 Hz), 144.8, 143.3, 132.4, 131.0 (t, 1C, J = 10.3 Hz), 120.0, 118.0, 114.2, 112.8 (t, 1C, J = 19.8 Hz), 112.1 (dd, 2C, J = 6.5 Hz, 19.1 Hz), 49.0, 33.5. APCI-MS m/z: 292.1 (MH⁺). HRMS calcd for C₁₅H₁₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0777.

Example 81 4,5-Dihydroxy-2-[(2-fluoro-3-chlorophenyl)methyl]-H-isoindol-l-one (24r).

[0173] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 21.9 minutes. ¹H NMR (DMSO): δ 7.46 (dt, IH, / = 1.6 Hz, 8.0 Hz), 7.21 (dt, IH, J = 1.6 Hz, 8.0 Hz), 7.15 (dt, IH, J = 0.8 Hz, 8.0 Hz), 6.91 (d, IH, J = 7.6 Hz), 6.72 (d, IH, J = 7.6 Hz), 4.69 (s, 2H), 4.19 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 155.8 (d, 1C, J = 246.4 Hz), 144.9, 143.4, 132.7, 130.2, 129.4 (d, 1C, J = 3.8 Hz), 126.7 (d, 1C, J = 14.6 Hz), 126.0 (d, 1C, J = 4.6 Hz), 120.2 (d, 1C, J = 17.5 Hz), 120.0, 118.0, 114.2, 49.3, 39.7. APCI-MS m/z: 308.0 (M⁺). HRMS calcd for Ci₅H₁₂NO₃FCl [MH⁺]: 308.0490. Found: 308.0489.

Example 82 2-(2-chloro-4-fluorobenzyl)-4,5-dihydroxyisoindoI-l-one (24s).

[0174] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 22.2 minutes. ¹H NMR (DMSO): δ 7.41 (dd, IH, J= 2.4 Hz, 8.8 Hz), 7.27 (dd, IH, 7 = 6.8 Hz, 8.4 Hz), 7.16 (dt, IH, J = 2.4 Hz, 8.8 Hz), 6.92 (d, IH, 7 = 8.0 Hz), 6.72 (d, IH, J = 8.0 Hz), 4.66 (s, 2H), 4.16 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 161.7 (d, 1C, 7 = 245.7 Hz), 144.9, 143.4, 133.6 (d, 1C, 7 = 10.6 Hz), 132.7, 131.6 (d, 1C, 7 = 9.1 Hz), 131.5 (d, 1C, 7 = 3.8 Hz), 120.0, 118.1, 117.2 (d, 1C, 7 = 24.4 Hz), 115.1 (d, 1C, 7 = 11.4 Hz), 114.2, 49.3, 43.0. APCI-MS m/z: 308.0 (MH⁺). HRMS calcd for C₁₅H₁₂NO₃FCl [MH⁺]: 308.0490. Found: 308.0489.

Example 83 2-(5-chloro-2-fluorobenzyl)-4,5-dihydroxyisoindol-l-one (24t)

[0175] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 25.9 minutes. ¹H NMR (DMSO): δ 7.37-7.33 (m, IH), 7.29 (dd, IH, 7 = 2.4 Hz, 6.4 Hz), 7.22 (t, IH, 7 = 9.2 Hz), 6.92 (d, IH, 7 = 8.0 Hz), 6.72 (d, IH, 7 = 8.0 Hz), 4.63 (s, 2H), 4.21 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 159.4 (d, 1C, 7 = 244.1 Hz), 144.9, 143.4, 132.7, 130.2 (d, 1C, 7 = 4.6 Hz), 129.8 (d, 1C, 7 = 4.6 Hz), 128.8 (d, 1C, 7 = 3.0 Hz), 126.9 (d, 1C, 7 = 16.8 Hz), 120.0, 118.0 (d, 1C, 7 = 6.1 Hz), 117.7, 114.2, 49.4, 39.5. APCI-MS m/z: 308.0 (MH⁺). HRMS calcd for C₁₅Hi₂NO₃FCl [MH⁺]: 308.0490. Found: 308.0489.

Example 84 2-(2-chloro-6-fluorobenzyl)-4,5-dihydroxyisoindol-l-one (24u).

[0176] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 20.2 minutes.

¹H NMR (DMSO): δ 7.39-7.33 (m, IH), 7.30 (d, IH, J = 8.0 Hz), 7.20 (t, IH, J = 8.8 Hz), 6.89 (d, IH, 7 = 8.0 Hz), 6.68 (d, IH, 7 = 8.0 Hz), 4.75 (d, 2H, J = 1.2 Hz), 4.04 (s, 2H). ¹³C NMR (DMSO): δ 167.8, 161.9 (d, 1C, 7 = 247.9 Hz), 144.8, 143.3, 135.2 (d, 1C, J = 5.3 Hz), 132.4, 131.2 (d, 1C, J = 9.1 Hz), 126.1 (d, 1C, J = 3.0 Hz), 122.5 (d, 1C, 7 = 7.5 Hz), 120.0, 118.0, 115.1 (d, 1C, 7= 22.9 Hz), 114.2, 48.8, 37.1 (d, 1C, 7 = 3.9 Hz). MALDI-MS m/z: 307.54 (MH⁺). HRMS calcd for C₁₅H]₂NO₃FCl [MH⁺]: 308.0490. Found: 308.0488.

Example 85 2-(3-fluoro-4-methylbenzyl)-4,5-dihydroxyisoindoI-l-one (24v).

[0177] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.3 minutes. ¹H NMR (DMSO): δ 7.18 (t, IH, 7 = 8.0 Hz), 6.98-6.90 (m, 3H), 6.70 (d, IH, 7 = 8.4 Hz), 4.56 (s, 2H), 4.14 (s, 2H), 2.14 (s, 3H). ¹³C NMR (DMSO): δ 168.4, 161.1 (d, 1C, 7 = 242.6 Hz), 144.8, 143.4, 138.0 (d, 1C, 7 = 6.9 Hz), 132.6, 132.2, 123.8 (d, 1C, 7 = 3.1 Hz), 123.5 (d, 1C, 7 = 17.5 Hz), 120.0, 118.2, 114.5 (d, 1C, 7 = 21.3 Hz), 114.2 (dd, 1C, 7 = 1.5 Hz, 5.3 Hz), 49.1, 44.9, 14.2 (t, 1C, 7 = 3.0 Hz). APCI-MS m/z: 288.0 (MH⁺). HRMS calcd for C₁₆Hi₅NO₃F [MH⁺]: 288.1036. Found: 288.1035.

Example 86 2-(4-fluoro-3-methylbeπzyI)-4,5-dihydroxyisoindol-l-one (24w).

[0178] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.2 minutes. ¹H NMR (DMSO): δ 9.30 (s, IH), 8.79 (s, IH), 7.13 (d, IH, J = 7.6 Hz), 7.09-7.02 (m, 2H), 6.90 (d, IH, 7 = 8.0 Hz), 6.70 (d, IH, 7 = 8.0 Hz), 4.53 (s, 2H), 4.13 (s, 2H), 2.15 (s, 3H). ¹³C NMR (DMSO): δ 168.3, 160.4(d, 1C, 7 = 241.1 Hz), 144.8, 143.4, 133.8 (d, 1C, 7 = 3.1 Hz), 132.6, 131.3 (d, 1C, J = 4.5 Hz), 127.4 (d, 1C, 7 = 8.3 Hz), 124.8 (d, 1C, J = 16.8 Hz), 119.9, 118.3, 115.4 (d, 1C, J = 22.9 Hz), 114.2, 49.0, 44.8, 14.5 (d, 1C, J = 3.9 Hz). HRMS calcd for C₁₆Hi₅NO₃F [MH⁺]: 288.1036. Found: 288.1035.

Example 87 4,5-dihydroxy-2-(perfluorobenzyl)isoindol-l-one (24x).

[0179] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 23.1 minutes. ¹H NMR (DMSO): δ 6.90 (dd, IH, J = 4.4 Hz, 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.73 (s, 2H), 4.20 (s, 2H). APCI-MS m/z: 346.0 (MH⁺). HRMS calcd for Ci₅H₉NO₃F₅ [MH⁺]: 346.0503. Found: 346.0500.

Example 88 2-(2,3-difluorobenzyl)-4,5-dihydroxyisoindol-l-one (24y).

[0180] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 19.4 minutes. ¹H NMR (DMSO): δ 7.33-7.28 (m, IH), 7.15-7.10 (m, IH), 7.05 (t, IH, J = 6.4 Hz), 6.92 (d, IH, J = 8.0 Hz), 6.72 (d, IH, J = 8.0 Hz), 4.69 (s, 2H), 4.19 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 150.1 (dd, IC, / = 12.2 Hz, 244.8 Hz), 148.4 (dd, 1C, J = 13.0 Hz, 245.6 Hz), 144.9, 143.4, 132.7, 127.3 (d, 1C, / = 12.2 Hz), 125.6 (t, 1C, J = 3.1 Hz), 125.4 (dd, 1C, / = 4.6 Hz, 6.9 Hz), 120.0, 118.0, 117.0 (d, 1C, / = 16.8 Hz), 114.2, 49.2, 39.2. APCI-MS m/z: 292.0 (MH⁺). HRMS calcd for C₁₅Hi₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0783.

Example 89 2-(2,4-difluorobeπzyl)-4,5-dihydroxyisoindol-l-one (24z).

[0181] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 18.6 minutes. ¹H NMR (DMSO): 5 7.31 (dd, IH, 7 = 8.4 Hz, 15.2 Hz), 7.22-7.16 (m, IH), 7.04-6.99 (m, IH), 6.91 (d, IH, J = 8.0 Hz), 6.71 (d, IH, 7= 8.0 Hz), 4.62 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 162.2 (dd, 1C, J = 12.2 Hz, 244.1 Hz), 160.7 (dd, 1C, J = 12.9 Hz, 247.1 Hz), 144.8, 143.4, 132.6, 131.9 (dd, 1C, J = 6.1 Hz, 10.0 Hz), 120.9 (dd, 1C, / = 3.8 Hz, 15.2 Hz), 120.0, 118.1, 114.2, 112.1 (dd, 1C, 7 = 3.8 Hz, 21.4 Hz), 104.4 (t, 1C, 7 = 25.9 Hz), 49.2, 39.1. APCI-MS m/z: 292.0 (MH⁺). HRMS calcd for C₁₅H₁₂NO₃F₂ [MH⁺]: 292.0785. Found: 292.0779. Example 90

2-(3-bromo-4-fluorobenzyl)-6,7-dihydroxyisoindolin-l-one (24aa).

[0182] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 24.0 minutes. ¹H NMR (DMSO): 5 9.31 (s, IH), 8.81 (s, IH), 7.56 (dd, IH, 7 = 2.0 Hz, 6.4 Hz), 7.33-7.24 (m, 2H), 6.91 (d, IH, 7 = 8.0 Hz), 6.71 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 4.58 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 157.9 (d, 1C, 7 = 243.3 Hz), 151.9, 148.4, 144.9, 143.4, 136.3 (d, 1C, 7 = 3.8 Hz), 133.0 (d, 1C, 7 = 7.7 Hz), 132.7, 120.0, 118.2, 117.3 (d, 1C, 7 = 22.1 Hz), 114.2, 108.5 (d, 1C, 7 = 21.3 Hz), 49.1, 44.4. APCI-MS m/z: 351.9 (MH⁺). HRMS calcd for Ci₅H₁₂NO₃FBr [MH⁺]: 351.9985. Found: 351.9984. Example 91

4,5-dihydroxy-2-(2,3,6-trifluorobenzyl)isoindol-l-one (24bb).

[0183] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 19.0 minutes. ¹H NMR (DMSO): δ 7.47-7.38 (m, IH), 7.13-7.07 (m, IH), 6.90 (d, IU, J = 8.0 Hz), 6.70 (d, IH, J = 8.0 Hz), 4.71 (s, 2H), 4.17 (s, 2H). APCI-MS m/z: 310.0 (MH⁺). HRMS calcd for Ci₅H_nNO₃F₃ [MH⁺]: 310.0691. Found: 310.0699.

Example 92 4,5-dihydroxy-2-(2,3,4-trifluorobenzyl)isoindoI-l-one (24cc).

[0184] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.2 minutes. ¹H NMR (DMSO): δ 7.23 (dd, IH, J = 8.4 Hz, 16.8 Hz), 7.12 (dd, IH, J = 6.4 Hz, 14.0 Hz), 6.91 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.66 (s, 2H), 4.19 (s, 2H). APCI-MS m/z: 310.0 (MH⁺). HRMS calcd for C₁₅Hi₁NO₃F₃ [MH⁺]: 310.0691. Found: 310.0690.

Example 93 4,5-dihydroxy-2-(2,4,5-trifluorobenzyl)isoindol-l-one (24dd).

[0185] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 20.8 minutes. ¹H NMR (DMSO): δ 7.52-7.45 (m, IH), 7.39-7.32 (m, IH), 6.91 (t, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.61 (s, 2H), 4.20 (s, 2H). APCI-MS m/z: 310.0 (MH⁺). HRMS calcd for C₁₅HnNO₃F₃ [MH⁺]: 310.0691. Found: 310.0693.

Example 94 4,5-dihydroxy-2-(3,4,5-trifluorobenzyl)isoindoI-l-one (24ee).

[0186] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.0 minutes. ¹H NMR (DMSO): δ 7.13 (t, 2H, / = 8.0 Hz), 6.92 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.57 (s, 2H), 4.19 (s, 2H). APCI-MS m/z: 310.0 (MH⁺). HRMS calcd for Ci₅HuNO₃F₃ [MH⁺]: 310.0691. Found: 310.0687.

Example 95 2-(2-chIoro-3,6-difluorobenzyl)-4,5-dihydroxyisoindol-l-one (24ff).

[0187] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 29.5 minutes. ¹H NMR (DMSO): δ 7.45-7.39 (m, IH), 7.31-7.25 (m, IH), 6.90 (d, IH, 7 = 7.6 Hz), 6.68 (d, IH, / = 7.6 Hz), 4.77 (s, 2H), 4.08 (s, 2H). ¹³C NMR (DMSO): δ 167.8, 157.6 (d, 1C, J = 244.1 Hz), 154.6 (d, 1C, J = 238.8 Hz), 144.8, 143.4, 132.5, 124.4 (d, 1C, J = 19.9 Hz), 122.0, 121.0, 117.9, 117.4 (dd, 1C, J = 9.9 Hz, 23.6 Hz), 115.9 (dd, 1C, J = 8.4 Hz, 25.2 Hz), 114.2, 48.8, 37.2. APCI-MS m/z: 326.0 (MH⁺). HRMS calcd for Ci₅H_nNO₃F₂Cl [MH⁺]: 326.0396. Found: 326.0396.

Example 97 2-(2,3-dihydro-lH-inden-2-yl)-4,5-dihydroxyisoindol-l-one (24hh).

[0188] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 20.6 minutes. ¹H NMR (DMSO): (57.21-7.19 (m, 2H), 7.14-7.11 (m, 2H), 6.90 (d, IH, J = 7.6 Hz), 6.69 (d, IH, / = 7.6 Hz), 4.98-4.95 (m, IH), 4.07 (s, 2H), 3.14 (dd, 2H, /= 8.0 Hz, 16.0 Hz), 2.99 (dd, 2H, J = 8.0 Hz, 16.0 Hz). ¹³C NMR (DMSO): <5 168.4, 144.6, 143.2 (2C), 141.3 (2C), 132.4, 127.0 (2C), 124.7 (2C), 120.0, 118.5, 114.2, 51.6, 46.2, 37.2 (2C). APCI-MS m/z: 282.0 (MH⁺). Example 98

2-(3-benzoylbenzyl)-4,5-dihydroxyisoindol-l-one (24ii).

Example 99 4,5-dihydroxy-2-(3-(3-(trifluoromethyl)-3H-diazirin-3-yl)benzyl)isoindol-l-one (24jj).

Example 100 4,5-dihydroxy-2-(pyridin-2-ylmethyl)isoindol-l-one (24kk).

Example 101 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindol-l-one (2411).

[0189] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 22.1 minutes. ¹H NMR (DMSO): δ 7.84-7.81 (m, 3H), 7.22 (s, IH), 7.46-7.41 (m, 2H), 7.35 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.92 (d, IH, J = 8.0 Hz), 6.69 (d, IH, J = 8.0 Hz), 4.76 (s, 2H), 4.17 (s, 2H). ¹³C NMR (DMSO): δ 168.5, 144.8, 143.5, 135.6, 133.4, 132.7 (2C), 128.8, 128.0 (2C), 126.7, 126.5 (2C), 126.3, 120.0, 118.3, 114.2, 49.2, 45.7. APCI-MS m/z: 306.1 (MH⁺). HRMS calcd for Ci₉Hi₆NO₃ [MH⁺]: 306.1130. Found: 306.1132.

Example 102 (S)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindol-l-one (24mm).

Example 103 (R)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yI)methyl)isoindoI-l-one (24nn).

Example 104

(S)-2-((2,3-dihydro-lH-inden-l-yl)methyl)-4,5-dihydroxyisoindol-l-one (24oo).

Example 105 (R)-2-((2,3-dihydro-lH-inden-l-yl)methyl)-4,5-dihydroxyisoindol-l-one (24pp).

Example 106 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindol-l-one (24qq).

Example 107 4,5-Dihydroxy-2-[(2-aminophenyl)methyl]-H-isoindol-l-one (24rr).

Example 108

N-(2-((4,5-dihydroxy-l-oxoisoindoIin-2-yl)methyl)phenyl)-N- (methylsulfonyl)methanesulfonamide (24ss).

Example 109 N-(2-((4,5-dihydroxy-l-oxoisoindolin-2-yl)methyl) phenyl)methanesulfonamide (24tt).

Examples 110-113

Example 110 General Procedure I for the Synthesis of Phthalimides 26a-e and 26g-i.

[0190] Triethylamine (2.0 mmol) was added dropwise to a solution of 3,4- dimethoxyphthalic anhydride 25 (1.0 mmol) (Baudart, M. G.; Hennequin, L. F. J. Antibiotics 1993, 46, 1458-1470.) and an appropriate amine (1.0 mmol) in toluene (5.0 mL) and the mixture was stirred at reflux (overnight). The solvent was evaporated and the residue was taken up in dichloromethane, dried (Na₂SO₄) and evaporated. The product was obtained following purification by silica gel column chromatography.

Example 111 4,5-Dimethoxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione (26a).

[0191] Treatment of benzylamine as described in general procedure I provided 26a in 16% yield. ¹H NMR (CDCl₃): δ 7.49 (dd, IH, 7 = 1.2 Hz, 8.0 Hz), 7.39 (dd, 2H, J = 1.2 Hz, 8.0 Hz), 7.29-7.23 (m, 3H), 7.05 (d, IH, J = 8.0 Hz), 4.76 (s, 2H), 4.10 (d, 3H, J = 1.2 Hz), 3.90 (d, 3H, J= 1.2 Hz); ¹³C NMR (CDCl₃): δ 167.3, 166.0, 157.7, 147.2, 136.5, 128.6 (4C), 127.7, 124.6, 121.9, 119.4, 115.7, 62.5, 56.6, 41.6; FAB-MS m/z 298.2 (MH⁺).

Example 112 4,5-Dimethoxy-2-[(2-phenyl)ethyl]-lH-isoindole-l,3(2H)-dione (26b).

[0192] Treatment of [(2-phenyl)ethyl] amine as described in general procedure I provided 26b in 61% yield. ¹H NMR (CDCl₃): δ 7.47 (d, IH, J = 8.0 Hz), 7.24-7.22 (m, 5H), 7.05 (d, IH, J = 8.0 Hz), 4.08 (s, 3H), 3.90 (s, 3H), 3.84 (t, 2H, J = 7.6 Hz), 2.93 (t, 2H, J = 7.6 Hz); ¹³C NMR (CDCl₃): δ 167.4, 166.1, 157.6, 147.1, 138.1, 128.8 (2C), 128.5 (2C), 126.5, 124.6, 121.8, 119.3, 115.6, 62.5, 56.6, 39.2, 34.5; FAB-MS m/z 312.1 (MH⁺). Example 113

4,5-Dimethoxy-2-[(l-naphthyl)methyl]-lH-isoindole-l,3(2H)-dione (26c).

[0193] Treatment of [(l-naphthyl)methyl]amine as described in general procedure I provided 26c in 18% yield. ¹H NMR (CDCl₃): «5 8.34 (d, IH, J = 8.8 Hz), 7.79 (dd, 2H, / = 8.0 Hz, 24.4 Hz), 7.58-7.52 (m, IH), 7.50 (d, IH, J = 8.0 Hz), 7.48-7.44 (m, 2H), 7.39 (t, IH, J = 8.0 Hz), 7.04 (d, IH, J = 8.0 Hz), 5.24 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H); ¹³C NMR (CDCl₃): S 167.5, 166.2, 157.7, 147.2, 133.7, 131.5, 131.2, 128.6, 128.5, 127.4, 126.4, 125.7, 125.3, 124.5, 123.5, 121.9, 119.5, 115.8, 62.5, 56.6, 39.5; FAB-MS m/z 348.1 (MH⁺).

Example 114

4,5-Dimethoxy-2-[(4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (26d).

[0194] Treatment of [(4-fluorophenyl)methyl] amine as described in general procedure I provided 26d in 52% yield. ¹H NMR (CDCl₃): δ 7.48 (d, IH, J = 8.0 Hz), 7.37 (dd, 2H, J = 5.2 Hz, 8.4 Hz), 7.05 (d, IH, 7 = 8.0 Hz), 6.94 (t, 2H, J = 8.4 Hz), 4.71 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H); ¹³C NMR (CDCl₃): δ 167.3, 166.0, 163.5, 161.1, 157.8, 147.3, 132.3 (d, 1C, J = 3.1 Hz), 130.5 (d, 1C, J = 8.4 Hz), 124.4, 121.9, 119.5, 115.8, 115.5, 115.3, 62.5, 56.6, 40.8; FAB-MS m/z 315.3 (MH⁺).

Example 115 4,5-Dimethoxy-2-[(3-chloro-4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (26e).

[0195] Treatment of [(3-chloro-4-fluorophenyl)methyl]amine as described in general procedure I provided 26e in 29% yield. ¹H NMR (CDCl₃): δ 7.50 (dd, IH, J = 0.4 Hz, 8.0 Hz), 7.44 (dd, IH, J = 2.0Hz, 6.8 Hz), 7.30-7.26 (m, IH), 7.06 (d, IH, J = 8.4 Hz), 7.02 (d, IH, / = 8.4 Hz), 4.69 (s, 2H), 4.10 (s, 3H), 3.91 (s, 3H); ¹³C NMR (CDCl₃): δ 167.1, 165.8, 158.9, 157.8, 156.4, 147.3, 133.5 (d, 1C, J = 3.9 Hz), 131.0, 128.6 (d, 1C, J = 6.9 Hz), 124.3, 121.7, 119.6, 116.6 (d, 1C, J = 21.4 Hz), 115.9, 62.5, 56.6, 40.4; FAB-MS m/z 350.0 (MH⁺).

Example 116 4,5-Dimethoxy-2-[(3-(phenylmethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-dione (26g).

[0196] Treatment of [(3-(phenylmethyl)phenyl)methyl]amine as described in general procedure I provided 26g in 52% yield. ¹H NMR (CDCl₃): δ 7.50-7.49 (m, IH), 7.26-7.20 (m, 5H), 7.18-7.13 (m, 3H), 7.05 (d, IH, J = 8.4 Hz), 7.04-7.02 (m, IH), 4.73 (s, 2H), 4.11 (s, 3H), 3.92 (s, 3H), 3.90 (s, 3H); ¹³C NMR (CDCl₃): δ 167.3, 166.0, 157.7, 147.2, 141.5, 140.8, 136.6, 129.2, 128.9 (2C), 128.7, 128.4 (2C), 128.3, 126.3, 126.0, 124.6, 121.9, 119.4, 115.7, 62.5, 56.6, 41.8, 41.5.

Example 117

4,5-Dimethoxy-2-[(3-fluorophenyI)methyl]-lH-isoindole-l,3(2H)-dione (26h).

[0197] Treatment of [(3-fluorophenyl)methyl]amine as described in general procedure I provided 26h in 58% yield. ¹H NMR (CDCl₃): δ 7.50 (dd, IH, J = 1.2 Hz, 8.0 Hz), 7.26-7.20 (m, IH), 7.16-7.09 (m, 2H), 7.06 (dd, IH, J = 1.2 Hz, 8.0 Hz), 6.93-6.88 (m, IH), 4.73 (s, 2H), 4.10 (s, 3H), 3.90 (s, 3H); FAB-MS m/z 315.3 (MH⁺). Example 118

4,5-Dimethoxy-2-[(3-(hydroxymethyl)phenyl)methyl]-lH-isoindoIe-l,3(2H)-dione (26i).

[0198] Treatment of [((3-hydroxymethyl)phenyl)methyl]amine as described in general procedure I provided 26i in 44% yield. ¹H NMR (CDCl₃): δ 7.43 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.34 (s, IH), 7.29-7.19 (m, 3H), 7.20 (d, IH, 7 = 8.0 Hz), 4.72 (s, 2H), 4.59 (s, 2H), 4.07 (d, 3H, 7 = 1.2 Hz), 3.88 (d, 3H, J = 1.2 Hz); ¹³C NMR (CDCl₃): δ 167.4, 166.0, 157.7, 147.2, 141.5, 136.7, 128.8, 127.7, 127.0, 126.3, 124.4, 121.8, 119.4, 115.8, 64.9, 62.5, 56.6, 41.4; FAB-MS m/z 328.1 (MH⁺). Example 119-161

[0199] The following were prepared by demethylation of intermediates 26 using General Procedure G.

Example 119 4,5-Hydroxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione (27a).

[0200] Treatment of 26a with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 30% B to 55% B over 30 minutes; retention time = 24.2 minutes) afford product 27a as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.26-7.16 (m, 5H), 7.14 (d, IH, 7 = 8.0 Hz), 6.97 (d, IH, 7 = 8.0 Hz), 4.81 (s, 2H), 4.68 (s, 2H); ¹³C NMR (DMSO): δ 168.1, 167.6, 152.3, 144.0, 137.0 (2C), 128.1 (2C), 127.5 (2C), 127.1, 122.4, 118.5, 115.9, 40.5; FAB-MS m/z 268.1 (M-H).

Example 120 4,5-Dihydroxy-2-[(2-phenyl)ethyl]-lH-isoindoIe-l,3(2H)-dione (27b).

[0201] Treatment of 26b with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 50% B over 30 minutes; retention time = 18.3 minutes) afford product 27b as a white solid following lyophilization. ¹H NMR (CD₃OD): δ 7.22-7.11 (m, 6H), 6.96 (d, IH, J = 7.6 Hz), 6.70 (d, IH, J = 8.0 Hz), 3.76 (dd, 2H, J = 7.6 Hz, 8.4 Hz), 2.89 (t, 2H, J = 7.6 Hz); ¹³C NMR (CD₃OD): δ 168.2, 167.8, 152.2, 143.8, 138.3, 128.4 (2C), 128.0 (2C), 126.1, 122.4, 118.4, 115.7, 115.5, 38.5, 34.0; FAB-MS m/z 284.1 (MH⁺). Example 121

4,5-Dihydroxy-2-[(l-naphthyl)methyI]-lH-isoindole-l,3(2H)-dione (27c).

[0202] Treatment of 26c with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 60% B over 30 minutes; retention time = 22.3 minutes) afford product 27c as a white solid following lyophilization. ¹H NMR (DMSO): δ 8.21 (d, IH, J = 8.0 Hz), 7.91 (d, IH, J = 8.0 Hz), 7.81 (d, IH, J = 8.0 Hz), 7.56- 7.51 (m, 3H), 7.39 (t, IH, J = 8.0 Hz), 7.26 (d, IH, J = 6.8 Hz), 7.17 (d, IH, J = 7.6 Hz), 7.04 (d, IH, 7 = 7.6 Hz), 5.10 (s, 2H); ¹³C NMR (DMSO): δ 167.9, 167.0, 153.0, 144.7, 133.7, 132.5, 130.8, 129.0, 128.2, 126.9, 126.4, 125.8, 125.2, 123.5, 122.6, 119.2, 116.3, 116.1, 38.9.FAB-MS m/z 320.1 (MH⁺).

Example 122 4,5-Dihydroxy-2-[(4-fluorophenyl)methyI]-lH-isoindole-l,3(2H)-dione (27d).

[0203] Treatment of 26d with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 30% B to 50% B over 30 minutes; retention time = 24.0 minutes) afford product 27d as a white solid following lyophilization. ¹H NMR (DMSO): δ 126 (dd, 2H, J = 6.0 Hz, 8.4 Hz), 7.13 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.08 (t, 2H, J = 8.4 Hz), 7.01 (dd, IH, 7 = 1.6 Hz, 8.0 Hz), 4.60 (s, 2H); ¹³C NMR (DMSO): δ 167.6, 166.8, 163.0, 160.6, 152.9, 144.7, 133.8 (d, 1C, J = 3.0 Hz), 129.9 (d, 1C, J = 8.4 Hz), 122.6, 119.2, 1 16.2, 116.1, 115.8, 115.6, 40.1; FAB-MS m/z 288.1 (MH⁺).

Example 123 4,5-Dihydroxy-2-[(3-chloro-4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27e).

[0204] Treatment of 26e with boron tribromide as described in general method G followed by preparative HPLC [YMC] (linear gradient of 40% B to 70% B over 30 minutes; retention time = 18.3 minutes) afford product 27e as a white solid following lyophilization. ¹H NMR (CD₃OD): (57.43 (dd, IH, J = 2.4 Hz, 7.2 Hz), 7.30-7.26 (m, IH), 7.19 (d, IH, 7 = 7.6 Hz), 7.14 (t, IH, 7 = 8.4 Hz), 6.99 (d, IH, 7= 7.6 Hz), 4.68 (s, 2H); ¹³C NMR (CD₃OD): δ 167.8, 167.4, 158.5, 156.1, 152.5, 144.1, 134.6, 130.1, 128.1 (d, 1C, 7= 7.6 Hz), 122.3, 118.5, 116.3 (d, 1C, 7= 21.4 Hz), 116.0, 115.5, 39.4; FAB-MS m/z 344.0 (M+Na⁺).

Example 124 4,5-Dihydroxy-2-[(3-(bromomethyI)phenyl)methyl]-lH-isoindole-l,3(2H)-dione (27f)

and 4,5-Dihydroxy-2-[(3-(hydroxymethyl)phenyl)methyl]-lH-isoindoIe-l,3(2H)-dioπe

(27i).

[0205] Treatment of 26i with boron tribromide as described in general method G afforded a mixture of products 27f and 27i, which were separated as white solids by preparative HPLC [YMC] (linear gradient of 40% B to 50% B over 30 minutes: 27f (retention time = 23.2 minutes); ¹H NMR (DMSO): δ 10.83 (bs, IH), 10.13 (bs, IH), 7.28-7.26 (m, 3H), 7.16-7.14 (m, 2H), 7.01 (d, IH, J = 8.0 Hz), 4.62 (s, 2H), 4.61 (s, 2H); ¹³C NMR (DMSO): δ 167.7, 166.8, 152.9, 144.7, 138.8, 138.1, 129.4, 128.7, 128.4, 127.7, 122.6, 119.2, 116.2, 116.1, 34.7 (2C); FAB-MS m/z 359.9 (M-H). 27i (retention time = 11.1 minutes); ¹H NMR (DMSO): δ 10.82 (s, IH), 10.11 (bs, IH), 7.22 (t, IH, J = 1.6 Hz), 7.15 (t, 2H, J = 6.8 Hz), 7.14 (d, IH, J = 7.6 Hz), 7.08 (d, IH, J = 7.6 Hz), 7.01 (d, IH, / = 8.0 Hz), 4.61 (s, 2H), 4.40 (s, 2H); FAB- MS m/z 298 (M-H). Example 125

4,5-Dihydroxy-2-[(3-(phenylmethyl)phenyl)methyl]-lH-Isoindole-l,3(2H)-dione (27g).

[0206] Treatment of 26g with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 50% B to 70% B over 30 minutes; retention time = 18.6 minutes) afford product 27g as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.22-7.11 (m, 8H), 7.05-7.00 (m, 3H), 4.58 (s, 2H), 3.85 (s, 2H); ¹³C NMR (DMSO): δ 167.7, 166.8, 152.9, 144.6, 141.9, 141.4, 137.7, 129.1 (4C), 128.8 (3C), 128.1, 126.4, 125.3, 122.7, 119.2, 116.2, 41.3, 40.8; FAB-MS m/z 360.1 (MH⁺).

Example 126 4,5-Dihydroxy-2-[(3-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27h).

[0207] Treatment of 26h with boron tribromide as described in general method G followed by preparative HPLC [Phe] (linear gradient of 30% B to 60% B over 30 minutes; retention time = 22.5 minutes) afford product 27h as a white solid following lyophilization. ¹H NMR (DMSO): δ 10.84 (bs, IH), 10.13 (bs, IH), 7.34-7.29 (m, IH), 7.14 (d, IH, 7 = 7.6 Hz), 7.06- 7.04 (m, 3H), 7.01 (d, IH, 7 = 7.6 Hz), 4.63 (s, 2H); ¹³C NMR (DMSO): δ 167.6, 166.8, 163.8, 161.4, 152.9, 144.7, 140.4 (d, 1C, J = 7.7 Hz), 131.0 (d, 1C, J = 8.4 Hz), 123.6, 122.6, 119.2, 116.2(d, 1C, 7 = 15.2 Hz), 114.6, 114.4, 40.1; FAB-MS m/z 288.0 (MH⁺).

Example 127 4,5-Dihydroxy-2-[(2-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27i).

[0208] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 18.6 minutes. ¹H NMR (DMSO): δ 7.28-7.23 (m, IH), 7.20-7.16 (m, IH), 7.14 (d, IH, 7 = 8.0 Hz), 7.14- 7.06 (m, 2H), 7.02 (d, IH, J = 8.0 Hz), 4.67 (s, 2H). ¹³C NMR (DMSO): δ 167.5, 166.7, 160.2 (d, 1C, 7 = 244. I Hz), 152.9, 144.7, 129.9 (d, 1C, 7 = 7.6 Hz), 129.8 (d, 1C, 7 = 3.8 Hz), 124.9 (d, 1C, 7 = 3.1 Hz), 124.2 (d, 1C, 7 = 14.5 Hz), 122.6, 119.2, 116.2, 115.8 (d, 2C, 7 = 20.6 Hz), 34.9 (d, 1C, 7 = 4.6 Hz). APCI-MS m/z: 288.1 (MH⁺). HRMS calcd for C]₅H_{1 1}NO₄F [MH+]: 288.0672. Found: 288.0664.

Example 128 4,5-Dihydroxy-2-[(3-(azidomethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-dione (27j)

[0209] Sodium azide (29 mg, 0.45 mmol) was added to the solution of 151D-68 (80 mg, 0.22 mmol) in acetone-water (5/1, 2 mL). The mixture was refluxed overnight and purified by prepared HPLC. Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.5 minutes. White solid (15 mg, 21% yield) was afforded.Η NMR (CD₃OD): δ 7.30-7.27 (m, 3H), 7.21-6.19 (m, IH), 7.17 (d, IH, 7 = 8.0 Hz), 6.98 (d, IH, 7= 8.0 Hz), 4.72 (s, 2H), 4.29 (s, 2H). ¹³C NMR (CD₃OD): δ 168.0, 167.6, 152.4, 144.0, 137.7, 136.2, 128.6, 127.4, 127.3, 127.1, 122.4, 118.5, 115.9, 115.6, 53.9, 40.4. APCI-ESI m/z: 347.1 (MNa⁺). MALDI- MS m/z: 346.72 (MNa⁺). HRMS calcd for C₁₆H_nN₄O₄ [M-H]: 323.0780. Found: 323.0778. Example 129

4,5-Dihydroxy-2-[(3-chlorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27k).

[0210] Linear gradient of 30% B to 60% B over 30 minutes; retention time = 27.1 minutes. ¹H NMR (DMSO): δ 7.32-7.30 (m, IH), 7.28-7.25 (m, 2H), 7.18-7.17 (m, IH), 7.15 (dd, IH, J - 0.8 Hz, 8.0 Hz), 7.02 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 4.62 (s, 2H). ¹³C NMR (DMSO): δ 167.7, 166.8, 153.0, 144.7, 140.0, 133.5, 130.9, 127.7, 127.6, 126.4, 122.6, 119.2, 116.3, 116.1, 40.3. MALDI-MS m/z: 303.54 (M⁺). HRMS calcd for C₁₅H₁₁NO₄Cl [MH⁺]: 304.0377. Found: 304.0387.

Example 130 4,5-Dihydroxy-2-[(3-bromophenyl)methyl]-lH-isoindole-l,3(2H)-dione (271).

[0211] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 25.2 minutes. ¹H NMR (DMSO): δ 7.42 (d, IH, / = 1.6 Hz), 7.40 (t, IH, J = 2.0 Hz), 7.23 (dd, 2H, J = 7.6 Hz, 14.8 Hz), 7.15 (d, IH, J = 7.6 Hz), 7.02 (d, IH, J = 7.6 Hz), 4.62 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.8, 153.0, 144.7, 140.3, 131.2, 130.6, 130.5, 126.8, 122.6, 122.1, 119.2, 116.3, 116.1, 40.2. MALDI-MS m/z: 347.45 (M⁺). HRMS calcd for Ci₅H_nNO₄Br [MH⁺]: 347.9871. Found: 347.9862.

Example 131 4,5-Dihydroxy-2-[(3-iodophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27m).

[0212] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 23.9 minutes. ¹H NMR (DMSO): δ 7.60 (d, IH, / = 1.6 Hz), 7.58 (d, IH, J = 7.6 Hz), 7.22 (d, IH, J = 7.6 Hz), 7.14 (d, IH, J = 7.6 Hz), 7.08 (t, IH, J = 7.6 Hz), 7.02 (d, IH, J = 7.6 Hz), 4.59 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.8, 153.0, 144.7, 140.2, 136.5, 136.4, 131.2, 127.2, 122.6, 119.2, 116.3, 116.1, 95.3, 40.1. MALDI-MS m/z: 395.42 (M⁺). HRMS calcd for Ci₅H_nNO₄I [MH⁺]: 395.9733. Found: 395.9722.

Example 132 4,5-Dihydroxy-2-[(3, 4-difluorophenyl)methyl] -lH-isoindole-l,3(2H)-dione (27n).

[0213] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.2 minutes. ¹H NMR (DMSO): δ 7.34-7.25 (m, 2H), 7.14 (d, IH, 7 = 8.0 Hz), 7.07-7.04 (m, IH), 7.02 (d, IH, 7 = 8.0 Hz), 4.61 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.7, 152.9, 149.7 (dd, 1C, J = 13.0 Hz, 244.8 Hz), 149.1 (dd, 1C, J = 12.2 Hz, 243.4 Hz), 144.7, 135.3 (dd, 1C, J = 3.9 Hz, 5.3 Hz), 124.5 (dd, 1C, 7 = 3.8 Hz, 6.8 Hz), 122.6, 119.2, 118.4 (dd, 1C, 7 = 17.5 Hz, 96.1 Hz), 118.0, 117.0 (dd, 1C, 7 = 17.5 Hz, 80.1 Hz), 116.2, 55.3. MALDI-MS mJz: 305.66 (MH⁺). HRMS calcd for C]₅H₈NO₄F₂ [M-H]: 304.0421. Found: 304.0425.

Example 133 4,5-Dihydroxy-2-[(3,5-difluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27o).

[0214] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.1 minutes. ¹H NMR (DMSO): δ 7.15 (d, IH, 7= 7.6 Hz), 7.07 (tt, IH, 7= 2.4 Hz, 9.2 Hz), 7.02 (d, IH, 7 = 7.6 Hz), 6.95-6.91 (m, 2H), 4.65 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.8, 162.9 (d, 1C, 7 = 244.9 Hz), 162.8 (d, 1C, 7 = 245.6 Hz), 152.9, 144.7, 142.1 (t, 1C, 7 = 9.1 Hz), 122.6, 119.2, 116.2, 116.1, 110.8 (dd, 2C, J = 6.9 Hz, 18.3 Hz), 103.2 (t, 1C, 7 = 26.0 Hz), 40.1. MALDI-MS m/z: 305.67 (MH⁺). HRMS calcd for C₁₅H₁₀NO₄F₂ [MH⁺]: 306.0578. Found: 306.0585.

Example 134 4,5-Dihydroxy-2-[(2,5-difluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27p).

[0215] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 20.9 minutes.

¹H NMR (DMSO): δ 7.20 (dt, IH, J = 4.4 Hz, 9.2 Hz), 7.14 (d, IH, J = 7.6 Hz), 7.12-7.04 (m, 2H), 7.03 (d, IH, J = 7.6 Hz), 4.66 (s, 2H). ¹³C NMR (DMSO): δ 167.5, 166.6, 158.5 (d, 1C, J = 239.5 Hz), 156.4 (d, 1C, J = 240.3 Hz), 152.9, 144.7, 126.2 (dd, 1C, J = 8.4 Hz, 17.6 Hz), 122.6, 119.2, 117.3 (dd, 1C, / = 8.4 Hz, 23.7 Hz), 116.4 (dd, 1C, J = 4.5 Hz, 16.0 Hz), 116.3, 116.1, 116.1 (dd, 1C, J = 8.4 Hz, 13.0 Hz), 34.9. APCI-MS m/z: 306.0 (MH⁺). HRMS calcd for C₁₅H₁₀NO₄F₂ [MH⁺]: 306.0578. Found: 306.0571.

Example 135 4,5-Dihydroxy-2-[(2,6-difluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (27q).

[0216] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 17.1 minutes. ¹H NMR (DMSO): δ 7.37-7.30 (m, IH), 7.11 (d, IH, J = 8.0 Hz), 7.05-6.99 (m, 2H), 6.99 (d, IH, J = 8.0 Hz), 4.69 (s, 2H). ¹³C NMR (DMSO): δ 167.0, 166.2, 161.8 (d, 2C, J = 247.1 Hz), 152.9, 144.6, 130.5 (t, 1C, J = 10.3 Hz), 122.5, 119.2, 116.1, 116.1 (d, 1C, 7 = 13.7 Hz), 112.6, 111.9 (dd, 2C, /= 6.9 Hz, 18.3 Hz), 29.8. APCI-MS m/z: 306.0 (MH⁺). HRMS calcd for C₁₅H₈NO₄F₂ [M-H]: 304.0421. Found: 304.0422.

Example 136 4,5-Dihydroxy-2-[(2-fluoro-3-chlorophenyl)methyl]-lH-isoindole-l,3(2H)-dioπe (27r).

[0217] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 22.1 minutes. ¹H NMR (DMSO): δ 10.84 (brs, IH), 10.1 (brs, IH), 7.47-7.43 (m, IH), 7.21-7.17 (m, IH), 7.15 (d, IH, 7 = 7.6 Hz), 7.14-7.10 (m, IH), 7.02 (d, IH, / = 7.6 Hz), 4.70 (s, 2H). ¹³C NMR (DMSO): δ 167.4, 166.5, 155.4 (d, 1C, J= 247.2 Hz), 153.0, 144.7, 130.1, 128.9 (d, 1C, 7 = 3.8 Hz), 126.3 (d, 1C, 7 = 23.5 Hz), 125.8 (d, 1C, 7 = 4.6 Hz), 122.6, 120.1 (d, 1C, 7 = 16.8 Hz), 119.2, 116.2 (d, 1C, 7 = 22.1 Hz), 116.2, 34.9. MALDI-MS m/z: 321.52 (M⁺). HRMS calcd for C₁₅H₈NO₄FCl [M-H]: 320.0126. Found: 320.0121.

Example 137 2-(2-chloro-4-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27s).

[0218] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 24.0 minutes. ¹H NMR (DMSO): δ 10.86 (brs, IH), 10.15 (brs, IH), 7.41 (dd, IH, 7 = 2.4 Hz, 8.8 Hz), 7.20 (dd, IH, 7 = 7.6 Hz, 8.8 Hz), 7.16 (d, IH, 7 = 8.0 Hz), 7.11 (dt, IH, 7 = 2.4 Hz, 8.8 Hz), 7.03 (d, IH, 7 = 8.0 Hz), 4.66 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.7, 161.5 (d, 1C, 7 = 245.6 Hz), 153.0, 144.7, 132.8 (d, 1C, 7 = 10.7 Hz), 130.8 (d, 1C, 7 = 3.8 Hz), 130.5 (d, 1C, 7 = 9.1 Hz), 122.6, 119.2, 117.1 (d, 1C, 7 = 25.2 Hz), 116.3, 116.1, 114.9 (d, 1C, 7 = 20.6 Hz), 38.4. MALDI-MS m/z: 339.61 (MH⁺). HRMS calcd for C₁₅H₈NO₄FCl [M-H]: 320.0126. Found: 320.0127. Example 138

2-(5-chloro-2-fluorobenzyl)-4,S-dihydroxyisoindoline-l,3-dione (27t)

[0219] Linear gradient of 40% B to 45% B over 30 minutes; retention time = 26.3 minutes. ¹H NMR (DMSO): δ 7.35-7.31 (m, IH), 7.28 (dd, IH, 7 = 2.8 Hz, 6.4 Hz), 7.20 (t, IH, J = 9.6 Hz), 7.14 (d, IH, 7= 7.6 Hz), 7.02 (d, IH, J = 7.6 Hz), 4.66 (s, 2H). ¹³C NMR (DMSO): <5 167.4, 166.6, 159.0 (d, 1C, J = 244.9 Hz), 153.0, 144.7, 129.7, 129.7 (d, 1C, J = 12.2 Hz), 128.6 (d, 1C, J = 3.0 Hz), 126.4 (d, 1C, 7 = 16.8 Hz), 122.6, 119.2, 117.8 (d, 1C, J = 22.9 Hz), 116.3, 116.2, 34.8 (d, 1C, J = 3.8 Hz). MALDI-MS m/z: 321.69 (MH⁺). HRMS calcd for C₁₅H₈NO₄FCl [M-H]: 320.0126. Found: 320.0130.

Example 139 2-(2-chloro-6-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27u).

[0220] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 20.3 minutes. ¹H NMR (DMSO): δ 7.35-7.29 (m, IH), 7.25 (d, IH, 7 = 8.4 Hz), 7.15 (t, IH, J = 9.0 Hz), 7.10 (d, IH, J = 8.0 Hz), 6.99 (d, IH, 7 = 8.0 Hz), 4.75 (s, 2H). ¹³C NMR (DMSO): δ 167.0, 166.3, 161.8 (d, 1C, J = 248.7 Hz), 152.9, 144.5, 134.6 (d, 1C, J = 5.4 Hz), 130.6 (d, 1C, J = 9.9 Hz), 125.9 (d, 1C, 7 = 3.8 Hz), 122.5, 122.3 (d, 1C, 7 = 6.0 Hz), 119.2, 116.1, 116.0 (d, 1C, 7 = 15.3 Hz), 114.9 (d, 1C, 7 = 22.8 Hz), 33.5. MALDI-MS m/z: 321.50 (MH⁺). HRMS calcd for Ci₅H₈NO₄FCl [M-H]: 320.0126. Found: 320.0130.

Example 140 2-(3-fluoro-4-methylbenzyl)-4,5-dihydroxyisoindoline-l,3-dione illy).

[0221] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.8 minutes. ¹H NMR (DMSO): δ 7.15 (t, IH, 7 = 8.0 Hz), 7.13 (d, IH, J = 8.0 Hz), 7.01 (d, IH, J = 8.0 Hz), 6.98-6.92 (m, 2H), 4.58 (s, 2H), 2.12 (s, 3H). ¹³C NMR (DMSO): δ 167.6, 166.8, 160.9 (d, 1C, 7 = 241.9 Hz), 152.9, 144.7, 137.5 (d, 1C, 7 = 6.8 Hz), 132.1, 123.5 (d, 1C, 7 = 16.6 Hz), 123.4, 122.6, 119.2, 116.1, 116.1 (d, 1C, 7 = 10.7 Hz), 114.3 (d, 1C, 7 = 24.4 Hz), 40.1, 14.2. MALDI-MS m/v 301.57 (MH⁺). HRMS calcd for C₁₆HnNO₄F [M-H]: 300.0672. Found: 300.0682.

Example 141 2-(4-fluoro-3-methylbenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27w).

[0222] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 22.9 minutes. ¹H NMR (DMSO): 5 7.15-7.11 (m, IH), 7.13 (d, IH, 7 = 8.0 Hz), 7.08-7.04 (m, IH), 7.03- 6.98 (m, IH), 7.01 (d, IH, 7 = 8.0 Hz), 4.56 (s, 2H), 2.13 (d, 3H, 7 = 2.0 Hz). ¹³C NMR (DMSO): δ 167.6, 166.8, 160.3 (d, 1C, 7 = 240.3 Hz), 152.9, 144.7, 133.5 (d, 1C, 7 = 3.8 Hz), 131.1 (t, 1C, 7 = 6.1 Hz), 127.2 (d, 1C, 7 = 8.4 Hz), 124.7 (d, 1C, 7 = 17.6 Hz), 122.6, 119.2, 116.1 (2C), 115.3 (d, 1C, 7 = 22.1 Hz), 40.1, 14.4. MALDI-MS m/z: 301.58 (MH⁺). HRMS calcd for Ci₆H₁₁NO₄F [M-H]: 300.0672. Found: 300.0680.

Example 142 4,5-dihydroxy-2-(perfluorobenzyl)isoindoline-l,3-dione (27x).

[0223] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 24.4 minutes. ¹H NMR (DMSO): δ 7.10 (d, IH, J = 8.0 Hz), 6.99 (d, IH, J = 8.0 Hz), 4.72 (s, 2H). MAIDI- MS m/z: 359.56 (MH⁺). HRMS calcd for C₁₅H₅NO₄F₅ [M-H]: 358.0139. Found: 358.0145.

Example 143 2-(2,3-difluorobenzyl)-4,5-dihydroxyisoindoIine-l,3-dione (27y).

[0224] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 18.9 minutes ¹H NMR (DMSO): δ 7.28 (dd, IH, J = 8.0 Hz, 16.8 Hz), 7.14 (d, IH, J = 1.6 Hz), 7.10 (dd, IH, J = 8.0 Hz, 13.2 Hz), 7.02 (d, 2H, J = 8.0 Hz), 4.71 (s, 2H). ¹³C NMR (DMSO): δ 167.4, 166.5, 153.0, 150.0 (dd, 1C, 7 = 12.6 Hz, 244.1 Hz), 148.1 (dd, 1C, J = 13.0 Hz, 246.4 Hz), 144.7, 126.8 (d, 1C, J = 11.4 Hz), 125.2 (dd, 1C, J = 4.6 Hz, 7.6 Hz), 125.0 (2C), 122.5, 119.2, 116.5 (dd, 1C, J = 16.8 Hz, 74.8 Hz), 116.3, 34.5. APCI-MS m/z: 304.1 (M-H). HRMS calcd for C₁₅H₈NO₄F₂ [M-H]: 304.0421. Found: 304.0421. Example 144

2-(2,4-difluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27z).

[0225] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 18.6 minutes. ¹H NMR (DMSO): δ 10.8(brs, IH), 10.1 (brs, IH), 7.31-7.25 (m, IH), 7.21-7.18 (m, IH), 7.14 (dt, IH, J = 2.0 Hz, 8.0 Hz), 7.01 (dt, IH, J = 2.0 Hz, 8.0 Hz), 6.99-6.97 (m, IH), 4.64 (s, 2H). ¹³C NMR (DMSO): δ 167.5, 166.6, 160.5 (d, 1C, J = 257.1 Hz), 160.3 (d, 1C, J = 250.4 Hz), 152.9, 144.7, 131.4 (dd, 1C, 7 = 5.3 Hz, 9.9 Hz), 122.6, 120.6 (d, 1C, J= 11.4 Hz), 119.2, 116.2, 116.1, 111.9 (dd, 1C, 7= 3.8 Hz, 21.3 Hz), 104.3 (t, 1C, 7= 26.0 Hz), 34.4. MALDI-MS m/z: 305.59 (MH⁺). HRMS calcd for C₁₅H₈NO₄F₂ [M-H]: 304.0421. Found: 304.0423.

Example 145 2-(3-bromo-4-fluorobenzyl)-4,5-dihydroxyisoindoIine-l,3-dione (27aa).

[0226] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 25.6 minutes. ¹H NMR (DMSO): δ 7.56 (d, IH, 7 = 6.8 Hz), 77.26 (d, 2H, 7 = 2.4 Hz), 7.14 (d, IH, 7 = 7.6 Hz), 7.01 (d, IH, 7 = 7.6 Hz), 4.61 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.8, 157.9 (d, 1C, 7 = 243.4 Hz), 153.0, 144.7, 135.8 (d, 1C, 7 = 3.0 Hz), 132.8 (d, 1C, 7 = 8.4 Hz), 129.3, 122.6, 119.2, 117.2 (d, 1C, 7 = 21.3 Hz), 116.2 (d, 1C, J = 17.5 Hz), 108.3 (d, 1C, 7 = 21.4 Hz), 39.7. MALDI-MS m/z: 367 ^'.46 (MH⁺). HRMS calcd for C₅H₈NO₄FBr [M-H]: 363.9621. Found: 363.9627.

Example 146 4,5-dihydroxy-2-(2,3,6-trifluorobenzyl)isoindoline-l,3-dione (27bb).

[0227] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 19.4 minutes. ¹H NMR (DMSO): δ 7.41-7.37 (m, IH), 7.11 (d, IH, 7 = 8.0 Hz), 7.08-7.03 (m, IH), 7.00 (d, IH, J = 8.0 Hz), 4.72 (s, 2H). APCI-MS m/z: 322.1 (M-H). HRMS calcd for C₁₅H₇NO₄F₃ [M- H]: 322.0327. Found: 322.0322.

Example 147 4,5-dihydroxy-2-(2,3,4-trifluorobenzyl)isoindoline-l,3-dione (27cc).

[0228] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.8 minutes. ¹H NMR (DMSO): δ 10.8 (brs, IH), 10.1 (brs, IH), 7.23-7.17 (m, IH), 7.14 (d, IH, 7 = 8.0 Hz), 7.14-7.07 (m, IH), 7.01 (d, IH, / = 8.0 Hz), 4.67 (s, 2H). APCI-MS m/z: (M-H) 322.1. HRMS calcd for C₁₅H₇NO₄F₃ [M-H]: 322.0327. Found: 322.0328. Example 148

4,5-dihydroxy-2-(2,4,5-trifluorobenzyl)isoindolin-l,3-dione (27dd).

[0229] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 21.9 minutes. ¹H NMR (DMSO): δ 7.52-7.45 (m, IH), 7.40-7.34 (m, IH), 7.17 (d, IH, J = 8.0 Hz), 6.86 (d, IH, J = 8.0 Hz), 4.65 (s, 2H). APCI-MS m/z: 322.1 (M-H). HRMS calcd for C]₅H₇NO₄F₃ [M- H]: 322.0327. Found: 322.0338.

Example 149 4,5-dihydroxy-2-(3,4,5-trifluorobenzyl)isoindoline-l,3-dione (27ee).

[0230] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 23.8 minutes. ¹H NMR (DMSO): δ 7.15 (t, 2H, J = 8.0 Hz), 7.13 (d, IH, J = 7.6 Hz), 7.01 (d, IH, J = 7.6 Hz), 4.60 (s, 2H). MALDI-MS m/z: 323.54 (MH⁺). HRMS calcd for C₁₅H₇NO₄F₃ [M-H]: 322.0327. Found: 322.0323.

Example 150 2-(2-chloro-3,6-difluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27ff).

[0231] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 19.2 minutes. ¹H NMR (DMSO): δ 10.8 (brs, IH), 10.1 (brs, IH), 7.39-7.34 (m, IH), 7.25-7.19 (m, IH), 7.10 (dd, IH, J = 5.2 Hz, 8.0 Hz), 7.00 (dd, IH, J = 5.2 Hz, 8.0 Hz), 4.77 (d, 2H, J = 3.23 Hz). ¹³C NMR (DMSO): δ 167.0, 166.2, 157.6 (d, 1C, / = 242.4 Hz), 154.4 (d, 1C, J = 241.1 Hz), 152.9, 144.6, 124.1 (d, 1C, J = 18.3 Hz), 122.4, 121.4 (dd, 1C, / = 6.1 Hz, 19.1 Hz), 119.2, 116.9 (dd, 1C, / = 9.2 Hz, 22.9 Hz), 116.2, 115.9, 115.6 (dd, 1C, J = 7.6 Hz, 25.2 Hz), 33.6. MALDI-MS m/z: 339.48 (MH⁺). HRMS calcd for C₁₅H₇NO₄F₂Cl [MH⁺]: 338.0032. Found: 338.0033.

Example 152 2-(2,3-dihydro-lH-inden-2-yl)-4,5-dihydroxyisoindoline-l,3-dione (27hh).

[0232] Linear gradient of 30% B to 55% B over 30 minutes; retention time = 29.5 minutes. ¹H NMR (DMSO)I (J T-IS-T-IS Cm, 2H), 7.13-7.11 (m, 2H), 7.11 (d, IH, J = 8.0 Hz), 7.01 (d, IH, J = 8.0 Hz), 4.91-4.87 (m, IH), 3.36 (dd, 2H, J = 9.2 Hz, 15.6 Hz), 3.08 (dd, 2H, J = 9.2 Hz, 15.6 Hz). ¹³C NMR (DMSO): δ 167.9, 167.1, 152.8, 144.5, 141.4 (2C), 126.9 (2C), 124.7 (2C), 122.7, 119.1, 116.1, 115.9, 49.0, 36.1 (2C). MALDI-MS m/z: 295.54 (MH⁺). HRMS calcd for C₁₇H₁₂NO₄ [M-H]: 294.0766. Found: 294.0769.

Example 153 2-(3-benzoylbenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27ii).

Example 154

4,5-dihydroxy-2-(3-(3-(trifluoromethyl)-3H-diazirin-3-yI)benzyl)isoindoIine-l,3-dione

(27jj).

Example 155 4,5-dihydroxy-2-(pyridin-2-ylmethyl)isoindoline-l,3-dione (27kk).

Example 156 4,5-dihydroxy-2-(naphthaIen-2-ylmethyl)isoindoline-l,3-dione (2711).

[0233] ¹H NMR (DMSO): δ 7.83-7.81 (m, 3H), 7.72 (s, IH), 7.44-7.38 (m, 3H), 7.17 (d, IH, J = 8.0 Hz), 7.04 (d, IH, J = 8.0 Hz), 4.80 (s, 2H). ¹³C NMR (DMSO): δ 167.8, 166.9, 152.9, 144.7, 135.2, 133.2, 132.6, 128.6, 128.0, 127.9, 126.7, 126.3, 126.1 (2C), 126.0, 122.7, 119.2, 116.2, 41.1. APCI-MS m/z: 318.1 (M-H). HRMS calcd for Ci₉H]₂NO₄ [M-H]: 318.0766. Found: 318.0759.

Example 157

(S)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindoline-l,3-dione (27mm).

Example 158

(R)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindoline-l,3-dione

(27nn).

Example 159

(S)-2-((2,3-dihydro-lH-inden-l-yl)methyl)-4,5-dihydroxyisoindoline-l,3-dione (27oo).

Example 160 (R)-2-((2,3-dihydro-lH-inden-l-yl)methyl)-4,5-dihydroxyisoindoline-l,3-dione (27pp).

Example 161 4,5-dihydroxy-2-(naphthaIen-2-ylmethyl)isoindoline-l,3-dione (27qq).

Example 162

2,3-Dihydro-4,5-dihydroxy-2-(4-fluorophenylmethyl)-H-isoindol-l-one (28).

[0234] Zinc dust (80 mg, 1.23 mmol) was added to a solution of 27d (59 mg, 0.21 mmol) in acetic acid (1.0 mL) at room temperature and the mixture was stirred (overnight). The resulting mixture was filtered and the filtrate was concentrated and the residue was purified by preparative HPLC [YMC] (linear gradient of 30% B to 50% B over 30 minutes; retention time = 16.4 minutes) to provide 28 as a white solid following lyophilization. ¹H NMR (CD₃OD): δ 7.29 (dd, 2H, J = 5.6 Hz, 8.8 Hz), 7.15 (d, IH, J = 8.0 Hz), 7.04 (t, 2H, J = 8.8 Hz), 6.87 (d, 2H, / = 8.0 Hz), 4.71 (s, 2H), 4.21 (s, 2H); FAB-MS m/z 274.1 (MH⁺). HRMS calcd for C₁₅H₁₃FNO₃ [MH+]: 274.0879. Found: 274.0888.

Example 163 4-Hydroxy-2-[(4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione (29).

[0235] To a suspension of 3-hydroxyphthalic anhydride (704 mg, 4.29 mmol) in anhydrous toluene (15.0 mL) was added dropwise [(4-fluorophenyl)methyl] amine (0.49 mL, 4.32 mmol) followed by triethylamine (1.20 mL, 8.63 mmol) and the mixture was stirred at reflux (overnight). The solvent was evaporated and the residue was crystallized from EtOAc to provide 29 as a yellow solid (133 mg, 11% yield). The solid was further purified by preparative HPLC [YMD] (linear gradient of 30% B to 65% B over 30 minutes; retention time = 24.6 minutes) to yield 29 as a white solid following lyophilization. ¹H NMR (DMSO): δ 7.56 (dd, IH, / = 7.2 Hz, 8.4 Hz), 7.30-7.23 (m, 3H), 7.16 (d, IH, J = 8.4 Hz), 7.10 (t, 2H, J = 8.8 Hz), 4.64 (s, 2H); FAB-MS m/z 269.9 (M-H). HRMS calcd for Ci₅H_nFNO₃ [MH+]: 272.0723. Found: 272.0728.

Example 164 4-Amino-6/7-dimethoxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione (30).

[0236] Treatment of 4-amino-6,7-dimethoxyphthalic anhydride (White, E. H.; Bursey, M. M. Analogs of ruminol. Synthesis and chemiluminescence of two methoxy-substituted aminophthalic hydrazides. J. Org. Chem. 1966, 31, 1912-1917) with benzyl amine as described in general procedure H, afforded bis-methyl ether 30 in 38% yield. ¹H NMR (CDCl₃): δ 7.39-7.36 (m, 2H), 7.28-7.20 (m, 3H), 6.20 (s, IH), 5.14 (bs, 2H), 4.71 (s, 2H), 3.94 (s, 3H), 3.79 (s, 3H); ¹³C NMR (CDCl₃): δ 168.8, 166.0, 159.5, 143.1, 139.9, 136.8, 128.5 (4C), 127.6, 121.7, 103.2, 102.4, 62.3, 56.3, 41.1; FAB-MS m/z 313.1 (MH⁺).

Example 165 4-Amino-6,7-dimethoxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione (32).

[0237] Compound 30 (74 mg, 0.179 mmol) and 2,3-dimethoxybenzoic acid chloride (36 mg, 0.179 mmol) was dissolved in dichloromethane (2 mL). Followed by triethylamine (50 mL) was added. The resultant mixture was stirred at r.t. for 2 days. Extracted with dichloromethane and dried by sodium sulfate. Filtered and solvent was evaporated. The residue was purified with silica gel column. Product 32 (42 mg, 58.9% yield) was afforded and the starting material 32 (23 mg, 68.9% conversion) was recovered. ¹H NMR (CDCl₃): δ 11.77 (s, IH), 8.80 (s, IH), 7.69 (dd, IH, J = 1.6 Hz, J = 8.0 Hz), 7.15-7.04 (m, 4H), 4.62 (s, 2H), 4.05 (s, 3H), 4.04 (s, 3H), 3.99 (s, 3H), 3.98 (s,3H), 3.87 (s, 3H), 3.82 (s, 3H); ¹³C NMR (CDCl₃): δ 164.9, 164.6, 163.4, 162.8, 159.4, 152.8, 152.3, 148.2, 148.0, 143.7, 135.3, 124.9 124.1, 123.8, 122.7, 121.6, 119.0, 118.1, 117.7, 117.4, 116.6, 109.3, 106.2, 62.5, 61.8, 60.3, 56.7 (2C), 56.1, 50.2; FAB-MS m/z 578.3 (MH⁺).

Example 166

4-Amino-6,7-dimethoxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione (33).

[0238] Following general procedure E and preparative YMC HPLC (linear gradient of 10% B to 50% B over 30 minutes; retention time = 26.5 minutes) afford product 33 as a white solid following lyophilization from compound 33. ¹H NMR (DMSO): δ 11.38 (bs, IH), 11.28 (s, IH), 10.54 (s, IH) 10.32 (bs, IH), 9.65 (bs, IH), 9.59 (bs, IH), 9.24 (bs, IH), 8.39 (s, IH), 7.34 (dd, IH, J = 1.6 Hz, J = 8.0 Hz), 7.05 (d, IH, J = 8.0 Hz), 6.93 (dd, IH, J = 1.6 Hz, / = 8.0 Hz), 6.82 (d, IH, J = 8.0 Hz), 6.73 (t, IH, J = 8.0 Hz), 4.48 (dd, 2H, J = 16.0 Hz, J = 23.2 Hz); ¹³C NMR (CDCl₃): δ 166.4, 165.4, 165.1, 163.5, 154.7, 146.5, 146.3, 145.4, 144.4, 142.1, 132.1, 131.6, 120.8 (2C), 119.4, 119.2, 115.5, 114.4, 112.5, 112.1, 105.3, 50.4; FAB-MS m/z 492.1 (MH⁺).

Example 167 4-Amino-6,7-dihydroxy-2-(phenyImethyl)-lH-isoindole-l,3(2H)-dione (31).

[0239] Treatment of 30 with boron tribromide as described in general method F followed by preparative HPLC [YMC] (linear gradient of 35% B to 45% B over 30 minutes; retention time = 15.8 minutes) afforded product 31 as a yellow solid following lyophilization.Η NMR (CD₃OD): δ 7.28-7.25 (m, 5H), 6.34 (s, IH), 4.67 (s, 2H); ¹³C NMR (CD₃OD): δ 168.5, 167.8, 154.4, 137.3, 128.1 (3C), 127.3 (3C), 126.9 (2C), 113.7, 106.5, 40.1; FAB-MS m/z 285.1 (MH⁺).

Example 168

N-(2-benzyl-6,7-dihydroxy-l,3-dioxoisoindolin-4-yl)-N- (methylsulfonyl)methanesulfonamide (34).

Example 169 4,5-dihydroxyisoindoline-l,3-dione (35).

1 -Benzyl-5-(4-fluoro-benzylcarbamoyl)-3,4-dihydroxy- lH-pyrroIe-2-carboxylic acid ethyl ester (36).

[0240] Trimethylaluminum (1.0 mL, 2.0 M in hexane, 2.0 mmol) was slowly added at r.t. to a solution of 4-fluorophenylamine (250 mg, 0.23 mmol) in dry dichloromethane (5.0 mL) under argon. The mixture was stirred at rt for 15 min. and diethyl l-benzyl-3,4-dihydroxy- lH-pyrrole-2,5-dicarboxylate (80 mg, 0.24 mmol) was added. The mixture was warmed to 40oC under argon until TLC indicated that the reaction had gone to completion. The reaction was carefully quenched with diluted HCl and extracted with dichloromethane. Organic phase was dried by sodium sulfate. The solvent was evaporated and the residue was purified by preparative HPLC [YMC] (linear gradient of 40% B to 70% B over 35 minutes; retention time = 31.6 minutes) which afforded product 36 as a white solid following lyophilization.Η NMR (CDCl₃): δ IAl (bs, IH), 7.21-7.13 (m, 5H), 6.94-6.90 (m, 4H), 5.94 (s, 2H), 4.45 (d, 2H, J = 5.6 Hz), 4.25 (dd, 2H, J = 7.2 Hz, 14.0 Hz), 1.19 (t, 3H, J = 7.2 Hz); ¹³C NMR (CDCl₃): δ 163.3, 163.0, 161.1, 160.8, 140.6, 139.3, 133.8 (2C), 132.7, 129.1 (2C), 128.3, 126.8, 125.9, 115.5, 115.3, 115.2, 108.5, 61.1, 48.9, 42.4, 14.1; FAB-MS m/z 413 (MH⁺).

Example 171

5-(4-Fluoro-benzylcarbamoyI)-3,4-dihydroxy-lH-pyrrole-2-carboxylic acid ethyl ester

(37).

[0241] The solution of 36 (28 mg, 0.068 mmol), anisole (1OmL, 0.092 mmol) and sulfuric acid (3mL) in trifluoroacetic acid (1.0 mL) was refluxed at 9O⁰C for 0.5 hour. After cooling to r.t., the residue was extracted with ethyl acetate and washed with saturated sodium bicarbonate solution. Organic phase was dried with sodium sulfate. Solvent was evaporated and the residue was purified by HPLC. .¹H NMR (CDCl₃): 3 8.66 (bs, IH), 7.27 (dd, 2H, / = 5.6 Hz, 8.8 Hz), 7.16 (bs, IH), 7.01-6.96 (m, 2H), 4.57 (d, 2H, 7 = 6.0 Hz), 4.34 (dd, 2H, J -- 12 Hz, 14.0 Hz), 1.34 (t, 3H, J = 7.2 Hz); FAB-MS m/z 323.1 (MH⁺)

Example 172

(N^t2E,N^l5E)-l-benzyl-N'²,N'⁵-bis(4-fluorobenzylidene)-3,4-dihydroxy-lH-pyrrole-2,5- dicarbohydrazide (38).

Example 173

(N'²E,N'⁵E)-l-benzyl-N'²,N^lS-bis(3-bromo-4-fluorobenzylidene)-3,4-dihydroxy-lH- pyrrole-2,5-dicarbohydrazide (39).

Example 174

(N^t2E,N^l5E)-l-benzyl-N^|2,N'⁵-bis(2,3-dihydroxybenzylidene)-3,4-dihydroxy-lH-pyrrole- 2,5-dicarbohydrazide (40).

Examples 175-207

[0242] The following were prepared by reacton with the appropriate amine using General Procedure H. Example 175

2-(3-chlorobenzyl)-6,7-dimethoxyisoindolin-l-one (23h)

[0243] Yield 63%. ¹H NMR (CDCl₃): δ 7.18 (d, IH, J = 1.6 Hz), 7.16-7.10 (m, 2H), 7.09- 7.07 (m, IH), 6.97 (d, IH, J = 8.0 Hz), 6.92 (d, IH, J = 8.0 Hz), 4.60 (s, 2H), 4.06 (s, 2H), 4.00 (s, 3H), 3.77 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 152.3, 147.2, 139.2, 134.5, 134.3, 130.0, 128.0, 127.7, 126.2, 124.4, 117.9, 116.6, 62.4, 56.7, 48.5, 45.7. APCI-MS m/z: 318.1 (MH⁺).

Example 176 2-(3-bromobenzyl)-6,7-dimethoxyisoindolin-l-one (23i)

[0244] Yield 58%. ¹H NMR (CDCl₃): δ 7.34 (t, IH, / = 1.6 Hz), 7.27 (dt, IH, 7 = 1.6 Hz, 8.0 Hz), 7.13 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.08 (t, IH, J = 7.6 Hz), 6.97 (d, IH, J = 8.0 Hz), 6.92 (d, IH, J = 8.0 Hz), 4.59 (s, 2H), 4.06 (s, 2H), 4.01 (s, 3H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.6, 152.3, 147.2, 139.5, 134.3, 131.0, 130.7, 130.3, 126.7, 124.4, 122.7, 117.9, 116.6, 62.4, 56.7, 48.5, 45.7. APCI-MS m/z: 362.0 (MH⁺).

Example 177 2-(3-iodobenzyl)-6,7-dimethoxyisoindolin-l-one (23j)

[0245] Yield 60%. ¹H NMR (CDCl₃): δ 7.54 (d, IH, J = 1.6 Hz), 7.46 (d, IH, J = 7.6 Hz), 7.16 (d, IH, J = 7.6 Hz), 6.96 (d, IH, J = 8.0 Hz), 6.92 (d, IH, J = 7.6 Hz), 6.90 (d, IH, J = 8.0 Hz), 4.56 (s, 2H), 4.05 (s, 2H), 4.00 (s, 3H), 3.77 (s, 3H). '³C NMR (CDCl₃): δ 166.6, 152.3, 147.2, 139.5, 136.9, 136.6, 134.3, 130.4, 127.4, 124.4, 117.9, 116.6, 94.6, 62.4, 56.7, 48.5, 45.5. APCI-MS m/z: 410.0 (MH⁺).

Example 178 2-(3,4-dichlorobenzyI)-6,7-dimethoxyisoindolin-l-one (23k)

[0246] Yield 53%. ¹H NMR (CDCl₃): δ 7.34 (s, IH), 7.33 (d, IH, J = 8.0 Hz), 7.10 (dd, IH, J = 2.0 Hz, 8.0 Hz), 7.03 (d, IH, J = 8.0 Hz), 6.97 (d, IH, / = 8.0 Hz), 4.63 (s, 2H), 4.13 (s, 2H), 4.05 (s, 3H), 3.84 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 152.4, 147.4, 137.5, 134.2, 132.7, 131.7, 130.7, 129.9, 127.5, 124.4, 117.9, 116.7, 62.5 (d, 1C, 7 = 2.2 Hz), 56.7 (d, 1C, J = 3.8 Hz), 48.5, 45.3.

Example 179 6,7-dimethoxy-2-(naphthalen-2-ylmethyl)isoindolin-l-one (231)

[0247] Followed the same procedure as 151D-103 from 2-(Bromomethyl)naphthalene, naphthalen-2-ylmethanarnine was afforded with 45% yield (two steps). ¹H NMR (CDCl₃): δ 7.80-7.77 (m, 3H), 7.70 (s, IH), 7.47-7.42 (m, 2H), 7.41-7.38 (m, IH), 3.99 (s, 2H). ¹³C NMR (CDCl₃): δ 140.5, 133.5, 132.5, 128.2, 127.7, 127.6, 126.1, 125.8, 125.5, 125.1, 46.5. APCI-MS m/z: 158.0 (MH⁺).

[0248] Yield 37%. ¹U NMR (CDCl₃): δ 7.72-7.69 (m, 3H), 7.66 (s, IH), 7.39-7.34 (m, 3H), 6.95 (d, IH, J = 8.4 Hz), 6.87 (d, IH, J = 8.4 Hz), 4.80 (s, 2H), 4.08 (s, 3H), 4.04 (s, 2H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 152.3, 147.2, 134.6, 134.5, 133.3, 132.8, 128.6, 127.7, 127.6, 126.9, 126.3, 126.2, 126.0, 124.7, 117.9, 116.4, 62.5, 56.7, 48.5, 46.4. APCI- MS m/z: 334.1 (MH⁺).

Example 180 (S)-6,7-dimethoxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindolin-l-one (23m)

[0249] Yield 63%. ¹H NMR (CDCl₃): δ 7.11-7.07 (m, 2H), 7.05-7.00 (m, 2H), 6.95-6.93 (m, 2H), 5.64 (t, IH, / = 7.4 Hz), 4.11 (d, IH, J = 16.8 Hz), 4.08 (s, 3H), 3.90 (d, IH, J = 16.8 Hz), 3.83 (s, 3H), 2.80-2.75 (m, 2H), 2.10-2.07 (m, IH), 1.99-1.95 (m, IH), 1.88-1.83 (m, 2H). ¹³C NMR (CDCl₃): δ 167.2, 152.3, 147.2, 138.0, 135.1, 134.7, 129.2, 127.5, 127.0, 126.2, 124.8, 117.8, 116.5, 62.5 (d, 1C, J = 3.0 Hz), 56.8 (d, 1C, J = 4.6 Hz), 49.6, 45.3, 29.4, 28.7, 21.7. α [MeOH, 22.0] = -109.7

Example 181 (R)-6,7 -dimethoxy -2- (1 ,2,3,4-tetrahydronaphthalen-l -yl)isoindolin- 1 -one (23n)

[0250] Yield 56%. ¹H NMR (CDCl₃): δ 7.11-7.06 (m, 2H), 7.05-7.00 (m, 2H), 6.96-6.94 (m, 2H), 5.65 (t, IH, 7 = 7.4 Hz), 4.12 (d, IH, J = 16.8 Hz), 4.09 (s, 3H), 3.91 (d, IH, 7 = 16.8 Hz), 3.84 (s, 3H), 2.86-2.73 (m, 2H), 2.12-2.06 (m, IH), 2.00-1.93 (m, IH), 1.91-1.82 (m, 2H). ¹³C NMR (CDCl₃): <5 167.2, 152.3, 147.2, 138.0, 135.1, 134.7, 129.2, 127.5, 127.0, 126.2, 124.9, 117.8, 116.5, 62.5 (d, 1C, 7 = 1.5 Hz), 56.8 (d, 1C, 7 = 2.8 Hz), 49.6, 45.3, 29.4, 28.7, 21.7. α [MeOH, 21.6] = 131.8

Example 182 (S)-2-(2,3-dihydro-lH-inden-l-yl)-6,7-dimethoxyisoindolin-l-one (23o)

[0251] Yield 55%. ¹H NMR (CDCl₃): δ 7.24-7.19 (m, 2H), 7.13-7.10 (m, 2H), 7.02 (d, IH, 7 = 8.0 Hz), 6.94 (d, IH, 7 = 8.0 Hz), 6.02 (t, IH, 7 = 7.6 Hz), 4.08 (s, 3H), 4.06 (d, IH, 7 = 16.8 Hz), 3.90 (d, IH, J = 16.8 Hz), 3.84 (s, 3H), 3.07-3.00 (m, IH), 2.96-2.88 (m, IH), 2.54- 2.45 (m, IH), 2.06-1.97 (m, 2H). ¹³C NMR (CDCl₃): δ 166.8, 152.3, 147.2, 143.5, 141.4, 134.6, 128.0, 126.8, 125.0, 124.9, 124.4, 117.8, 116.5, 62.5 (d, 1C, 7 = 3.8 Hz), 56.8 (d, 1C, 7 = 6.1 Hz), 56.1 (d, 1C, J = 5.4 Hz), 44.9 (t, 1C, J = 5.0 Hz), 30.6, 30.4. APCI-MS m/z: 310.1 (MH⁺). α [CHCl₃, 23] = -98.0.

Example 183 (R)-2-(2,3-dihydro-lH-inden-l-yl)-6,7-dimethoxyisoindolin-l-one (23p)

[0252] Yield 57%. ¹H NMR (CDCl₃): δ 7.24-7.16 (m, 2H), 7.13-7.07 (m, 2H), 7.01(d, IH, J= 8.0 Hz), 6.93 (d, IH, 7= 8.0 Hz), 6.01 (t, IH, J = 7.6 Hz), 4.07 (s, 3H), 4.05 (d, IH, 7 = 16.8 Hz), 3.89 (d, IH, 7 = 16.8 Hz), 3.83 (s, 3H), 3.06-2.87 (m, 2H), 2.52-2.44 (m, IH), 2.05- 1.96 (m, IH). ¹³C NMR (CDCl₃): δ 166.8, 152.3, 147.2, 143.5, 141.4, 134.6, 128.0, 126.7, 124.9 (2C), 124.4, 117.8, 116.5, 62.5 (d, 1C, 7 = 3.8 Hz), 56.8 (d, 1C, 7 = 5.4 Hz), 56.1 (d, 1C, J = 3.9 Hz), 44.9 (d, 1C, 7 = 3.8 Hz), 30.6, 30.4. APCI-MS m/z: 310.1 (MH⁺). α [CHCl₃, 23] = 91.5.

Example 184 2-(2,3-dihydro-lH-inden-2-yl)-6,7-dimethoxyisoindolin-l-one (23q)

[0253] Yield 54%. ¹H NMR (CDCl₃): δ 7.21-7.18 (m, 2H), 7.16-7.13 (m, 2H), 6.99 (d, IH, J = 8.4 Hz), 6.92 (d, IH, J = 8.4 Hz), 5.29-5.25 (m, IH), 4.03 (s, 3H), 3.98 (s, 2H), 3.81 (s, 3H), 3.27 (dd, 2H, J = 7.6 Hz, 16.4 Hz), 2.94 (dd, 2H, J = 5.2 Hz, 16.4 Hz). ¹³C NMR (CDCl₃): δ 166.4, 152.3, 147.1, 140.9 (2C), 134.3, 126.8 (2C), 124.9, 124.4 (2C), 117.8, 116.4, 62.5 (d, 1C, J = 1.5 Hz), 56.7 (d, 1C, J = 2.2 Hz), 51.4, 45.5, 37.7 (2C). APCI-MS m/z: 310.1 (MH⁺).

Example 185 2-(2-fluorobenzyI)-6,7-dimethoxyisoindolin-l-one (23r)

[0254] Yield 58%. ¹H NMR (CDCl₃): δ 7.30-7.26 (m, IH), 7.19-7.13 (m, IH), 7.02-6.92 (m, 4H), 4.71 (s, 2H), 4.14 (s, 2H), 4.02 (s, 3H), 3.79 (s, 3H). ¹³C NMR (CDCl₃): δ 166.6, 160.8 (d, 1C, J = 244.8 Hz), 152.2, 147.2, 134.5, 130.8 (d, 1C, J = 3.8 Hz), 129.4 (d, 1C, J = 7.6 Hz), 124.6, 124.5 (d, 1C, J = 7.6 Hz), 123.9 (d, 1C, J = 15.2 Hz), 117.8, 116.5, 115.3 (d, 1C, J = 21.4 Hz), 62.4, 56.7, 48.7, 39.5 (d, 1C, J = 3.8 Hz). APCI-MS m/z: 302.1 (MH⁺).

Example 186 2-(3-fluorobenzyl)-6,7-dimethoxyisoindoIin- 1 -one (23s)

[0255] Yield 54%. ¹H NMR (CDCl₃): δ 7.20-7.15 (m, IH), 6.98 (d, IH, J = 8.0 Hz), 6.97 (s, IH), 6.92 (d, IH, J = 8.0 Hz), 6.89-6.82 (m, 2H), 4.62 (s, 2H), 4.08 (s, 2H), 4.01 (s, 3H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 162.9 (d, 1C, J = 244.9 Hz), 152.3, 147.2, 139.7 (d, 1C, J = 6.9 Hz), 134.3, 130.2 (d, 1C, J = 8.4 Hz), 124.5, 123.6 (d, 1C, J = 2.3 Hz), 117.9, 116.7, 114.8 (d, 1C, J = 22.1 Hz), 114.4 (d, 1C, J = 21.4 Hz), 62.4, 56.6, 48.5, 45.7. APCI- MS m/z: 302.1 (MH⁺).

Example 187 2-(3-chloro-2-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23t)

[0256] Yield 52%. ¹H NMR (CDCl₃): δ 7.22-7.16 (m, 2H), 6.99 (d, IH, J = 8.0 Hz), 6.94 (d, IH, J = 8.0 Hz), 6.93-6.91 (m, IH), 4.71 (s, 2H), 4.16 (s, 2H), 3.99 (s, 3H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 156.2 (d, 1C, / = 247.2 Hz), 152.3, 147.2, 134.3, 129.9, 129.0 (d, 1C, J = 3.8 Hz), 125.8 (d, 1C, J = 14.5 Hz), 124.9 (d, 1C, J = 4.6 Hz), 124.3, 120.9 (d, 1C, J = 17.5 Hz), 117.9, 116.6, 62.4 (d, 1C, J = 3.0 Hz), 56.7 (d, 1C, J = 3.8 Hz), 48.8, 39.7 (d, 1C, J = 3.8 Hz). APCI-MS m/z: 336 A (M⁺).

Example 188 2-(4-chloro-3-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23u)

[0257] Yield 51%. ¹H NMR (CDCl₃): δ 7.26 (t, IH, J = 8.0 Hz), 7.04-7.02 (m, IH), 7.02 (d, IH, J = 8.0 Hz), 6.98-6.95 (m, IH), 6.96 (d, IH, J = 8.0 Hz), 4.63 (s, 2H), 4.12 (s, 2H), 4.03 (d, 3H, J = 1.6 Hz), 3.82 (d, 3H, J = 1.6 Hz). ¹³C NMR (CDCl₃): δ 166.7, 158.1 (d, 1C, J = 248.7 Hz), 152.4, 147.3, 138.2 (d, 1C, J = 6.1 Hz), 134.2, 130.8, 124.4 (d, 1C, 7 = 3.9 Hz), 124.3, 120.0 (d, 1C, J = 17.6 Hz), 117.9, 116.7, 116.2 (d, 1C, J = 20.6 Hz), 62.4 (d, 1C, J = 3.1 Hz), 56.7 (d, 1C, J = 3.0 Hz), 48.5, 45.4. APCI-MS m/z: 336.1 (MH⁺).

Example 189 2-(2-chIoro-4-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23v)

[0258] Yield 58%. ¹H NMR (CDCl₃): δ 7.29 (dd, IH, J = 6.4 Hz, 8.8 Hz), 7.04 (dd, IH, J = 2.8 Hz, 8.8 Hz), 7.01 (d, IH, 7 = 8.4 Hz), 6.97 (d, IH, J = 8.4 Hz), 6.86 (dd, IH, / = 2.8 Hz, 8.0 Hz), 4.76 (s, 2H), 4.16 (s, 2H), 4.02 (s, 3H), 3.81 (s, 3H). ¹³C NMR (CDCl₃): δ 166.8, 161.8 (d, 1C, 7 = 248.7 Hz), 152.3, 147.3, 134.4, 134.1 (d, 1C, 7 = 10.6 Hz), 131.5 (d, 1C, 7 = 8.4 Hz), 130.7 (d, 1C, J= 3.0 Hz), 124.4, 117.8, 116.9, 116.6 (d, 1C, J = 3.8 Hz), 114.5 (d, 1C, 7 = 20.6 Hz), 62.4 (d, 1C, J = 3.0 Hz), 56.7 (d, 1C, J = 3.8 Hz), 48.9, 43.0. APCI-MS m/z: 336.0 (MH⁺).

Example 190 2-(5-chloro-2-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23w)

[0259] Yield 58%. ¹H NMR (CDCl₃): δ 125-1.23 (m, IH), 7.11-7.08 (m, IH), 7.00-6.94 (m, IH), 6.93-6.88 (m, IH), 4.66 (s, 2H), 4.15 (s, 2H), 4.00 (d, 3H, J = 1.6 Hz), 3.78 (d, 3H, 7 = 1.6 Hz). ¹³C NMR (CDCl₃): δ 166.7, 159.3 (d, 1C, 7 = 244.8 Hz), 152.3, 147.2, 134.3, 130.3 (d, 1C, J = 3.8 Hz), 129.5 (d, 1C, J = 3.0 Hz), 129.3 (d, 1C, J = 8.4 Hz), 125.8 (d, 1C, 7 = 16.8 Hz), 124.3, 117.9, 116.7 (d, 1C, J = 23.7 Hz), 116.6, 62.4, 56.7, 48.8, 39.3 (d, 1C, J = 3.8 Hz). APCI-MS m/z: 336.1 (MH⁺).

Example 191 2-(2-chloro-6-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23x)

[0260] Yield 51%. ¹H NMR (CDCl₃): δ 7.23-7.13 (m, 2H), 6.99 (d, IH, J = 8.0 Hz), 6.98- 6.94 (m, IH), 6.92 (d, IH, J = 8.0 Hz), 4.89 (d, 2H, J = 2.0 Hz), 4.05 (s, 2H), 4.03 (s, 3H), 3.80 (s, 3H). ¹³C NMR (CDCl₃): δ 166.0, 161.8 (d, 1C, J = 249.5 Hz), 152.2, 147.3, 136.0 (d, 1C, 7 = 5.4 Hz), 134.4, 130.0 (d, 1C, 7 = 9.9 Hz), 125.6 (d, 1C, J = 3.1 Hz), 124.5, 122.3 (d, 1C, 7 = 17.6 Hz), 117.7, 116.4, 114.2 (d, 1C, 7 = 22.1 Hz), 62.5 (d, 1C, 7 = 2.3 Hz), 56.7 (d, 1C, 7 = 3.0 Hz), 48.1, 37.2 (d, 1C, 7 = 3.8 Hz). APCI-MS m/z: 336.0 (MH⁺).

Example 192 2-(4-fluoro-3-methyIbenzyl)-6,7-dimethoxyisoindolin-l-one (23y)

[0261] Yield 60%. ¹H NMR (CDCl₃): δ 7.05 (d, IH, J = 7.2 Hz), 7.02 (brs, IH), 6.98 (dd, IH, J = 1.6 Hz, 8.0 Hz), 6.93 (d, IH, 7 = 8.0 Hz), 6.84 (t, IH, 7 = 8.8 Hz), 4.58 (s, 2H), 4.07 (s, 2H), 4.03 (d, 3H, J = 1.6 Hz), 3.79 (d, 3H, J = 1.6 Hz). ¹³C NMR (CDCl₃): δ 166.5, 160.7 (d, 1C, /= 243.3 Hz), 152.3, 147.2, 134.4, 132.6 (d, 1C, / = 3.0 Hz), 131.3 (dd, 1C, / = 5.3 Hz, 15.3 Hz), 127.0 (dd, 1C, / = 9.2 Hz, 16.8 Hz), 125.1 (d, 1C, / = 17.6 Hz), 124.7, 117.8 (dd, 1C, / = 2.2 Hz, 4.5 Hz), 116.4 (dd, 1C, / = 3.1 Hz, 7.6 Hz), 115.0 (d, 1C, 7 = 22.1 Hz), 62.4 (dd, 1C, / = 9.1 Hz, 37.4 Hz), 56.7 (dd, 1C, / = 20.6 Hz, 41.2 Hz), 48.3 (t, 1C, / = 17.5 Hz), 45.5 (t, 1C, /= 9.2 Hz), 14.3 (dd, 1C, /= 3.8 Hz, 12.9 Hz). APCI-MS m/z: 316.1 (MH⁺).

Example 193 2-(3-fluoro-4-methylbenzyl)-6,7-dimethoxyisoindolin-l-one (23z)

[0262] Yield 68%. ¹H NMR (CDCl₃): δ 7.01 (t, IH, / = 8.0 Hz), 6.97 (d, IH, / = 8.4 Hz), 6.91 (d, IH, / = 8.4 Hz), 6.88-6.84 (m, 2H), 4.58 (s, 2H), 4.06 (s, 2H), 4.01 (s, 3H), 3.77 (s, 3H), 2.12 (s, 3H). ¹³C NMR (CDCl₃): δ 166.6, 161.3 (d, 1C, J = 245.1 Hz), 152.2, 147.2, 136.8 (d, 1C, / = 6.9 Hz), 134.4, 131.6 (dd, 1C, / = 5.3 Hz, 8.4 Hz), 124.6, 123.9 (d, 1C, 7 = 17.5 Hz), 123.4, 117.8 (dd, 1C, 7 = 2.3 Hz, 4.5 Hz), 116.5 (dd, 1C, / = 2.3 Hz, 6.1 Hz), 114.6 (dd, 1C, / = 2.3 Hz, 21.3 Hz), 62.4 (d, 1C, / = 18.3 Hz), 56.6 (d, 1C, /= 19.8 Hz), 48.4 (t, 1C, / = 17.6 Hz), 45.5 (dd, 1C, / = 8.4 Hz, 9.9 Hz), 14.1 (dd, 1C, / = 3.1 Hz, 12.2 Hz). APCI-MS m/z: 316.1 (MH⁺).

Example 194 2-(3-bromo-4-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23aa)

[0263] Borane-THF complex (1.0 M in THF, 17.3 niL, 17.3 mmol) was added dropwise to the solution of 3-bromo-4-fluorobenzonitrile (1.153 g, 5.8 mmol) in THF (anhydrous, 20 mL) carefully. The result solution was refluxed overnight, and then cooled to room temperature. Water was added dropwise carefully to quench the reaction. The mixture was extracted with chloroform and washed with brine. Organic phase was dried by anhydrous sodium sulfate and filtered. The solvent was concentrated and the residue was purified by silica gel column. (3- Bromo-4-fluorophenyl)methanamine as a colorless oil (571 mg) was afforded. Yield 49%. ¹H NMR (CDCl₃): δ 734 (d, IH, J = 4.4 Hz), 7.03 (t, IH, J = 2.0 Hz), 6.87 (dd, IH, J = 2.0 Hz, 4.0 Hz), 3.63 (d, 2H, 7 = 6.0 Hz), 1.46 (s, 2H). ¹³C NMR (CDCl₃): δ 157.6 (d, 1C, J = 244.1 Hz), 140.5 (d, 1C, J = 3.8 Hz), 131.9, 127.5 (d, 1C, 7 = 6.8 Hz), 116.1 (d, 1C, J = 12.1 Hz), 108.6 (d, 1C, J = 20.6 Hz), 45.0 (d, 1C, J = 1.6 Hz).

[0264] Yield 65%. ¹H NMR (CDCl₃): δ 7.43 (dd, IH, J = 1.6 Hz, 6.0 Hz), 7.19-7.15 (m, IH), 7.02 (dd, IH, J= 0.8 Hz, 8.0 Hz), 6.99 (s, IH), 6.96 (d, IH, 7= 8.0 Hz), 4.61 (s, 2H), 4.11 (s, 2H), 4.03 (s, 3H), 3.81 (s, 3H). ¹³C NMR (CDCl₃): S 166.7, 158.4 (d, 1C, 7 = 246.4 Hz), 152.4, 147.3, 134.7 (d, 1C, J = 3.7 Hz), 134.2, 133.0, 128.8 (d, 1C, 7 = 7.6 Hz), 124.4, 117.9, 116.7, 116.6 (d, 1C, 7 = 12.2 Hz), 109.2 (d, 1C, 7 = 21.4 Hz), 62.4, 56.7, 48.5, 45.1. APCI-MS m/z: 380.0 (MH⁺).

Example 195 2-(2,3-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23bb)

[0265] Hydrazine hydrate (3.0 mL, 61.6 mmol) was added to the suspension of 151D-88 (3.36 g, 12.3 mmol) in ethanol (50 mL). The result clear solution was refluxed for 2 hours. The solvent was vapored and the residue was washed with chloroform and filtered. The filtrate was concentrated and purified by silica gel column. (2,3-difluorophenyl)methanamine (1.13 g, 64% yield) was afforded. ¹H NMR (CDCl₃): δ 6.96-6.88 (m, 3H), 3.75 (s, 2H)). APCI-MS m/z: 144.0 (MH⁺). [0266] Yield 49%. ¹H NMR (CDCl₃): δ 7.05-7.01 (m, IH), 6.99 (d, IH, 7 = 8.0 Hz), 6.99- 6.97 (m, IH), 6.95 (d, IH, J = 8.0 Hz), 6.95-6.90 (m, IH), 4.72 (s, 2H), 4.15 (s, 2H), 3.99 (s, 3H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 152.3, 150.3 (dd, 1C, J = 12.9 Hz, 247.9 Hz), 148.6 (dd, 1C, / = 28.2 Hz, 259.4 Hz), 147.2, 134.3, 126.5 (d, 1C, J = 11.5 Hz), 125.3 (t, 1C, J = 3.0 Hz), 124.4, 124.4 (dd, 1C, J= 3.9 Hz, 9.9 Hz), 117.8, 116.6, 116.5 (d, 1C, 7 = 15.3 Hz), 62.4, 56.6 (d, 1C, J = 1.6 Hz), 48.8, 39.2. APCI-MS m/z: 320.1 (MH⁺).

Example 196 2-(2,4-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23cc)

[0267] Yield 61%. ¹H NMR (CDCl₃): δ 7.34-7.28 (m, IH), 7.01 (d, IH, 7 = 8.4 Hz), 6.96 (d, IH, 7 = 8.4 Hz), 6.78-6.71 (m, 2H), 4.68 (s, 2H), 4.16 (s, 2H), 4.02 (s, 3H), 3.81 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 162.4 (dd, 1C, 7 = 12.2 Hz, 247.5 Hz), 160.8 (dd, 1C, 7 = 12.2 Hz, 247.1 Hz), 152.3, 147.3, 134.4, 131.9 (dd, 1C, 7 = 5.3 Hz, 9.9 Hz), 124.5, 120.1 (dd, 1C, 7 = 3.8 Hz, 15.2 Hz), 117.8, 116.6, 111.7 (dd, 1C, 7 = 3.8 Hz, 21.3 Hz), 103.7 (t, 1C, 7 = 15.1 Hz), 62.4, 56.7, 48.7, 39.1(d, 1C, 7 = 3.1 Hz). APCI-MS m/z: 320.1 (MH⁺).

Example 197 2-(2,5-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23dd)

[0268] Yield 67%. ¹H NMR (CDCl₃): δ 6.98 (d, IH, 7 = 8.0 Hz), 6.94 (d, IH, 7 = 8.0 Hz), 6.96-6.88 (m, 2H), 6.83-6.81 (m, IH), 4.65 (s, 2H), 4.15 (s, 2H), 3.99 (s, 3H), 3.77 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 158.7 (d, 1C, 7 = 241.8 Hz), 156.7 (d, 1C, 7 = 242.5 Hz), 152.3, 147.2, 134.3, 125.8 (dd, 1C, 7 = 7.6 Hz, 17.5 Hz), 124.3, 117.8, 116.8 (dd, 1C, 7 = 8.4 Hz, 68.7 Hz), 115.9 (dd, 1C, 7 = 9.2 Hz, 67.9 Hz), 62.3, 56.6, 48.8, 39.4 (d, 1C, 7 = 3.8 Hz). APCI-MS m/z: 320.1 (MH⁺). Example 198 2-(3,4-difluorobenzyl)-6,7-dimethoxyisoindoIin-l-one (23ee)

[0269] Yield 58%. ¹H NMR (CDCl₃): δ 7.08-7.05 (m, IH), 7.04-6.99 (m, IH), 7.01 (d, IH, J = 8.0 Hz), 6.98-6.95 (m, IH), 6.96 (d, IH, J = 8.0 Hz), 4.61 (s, 2H), 4.11 (s, 2H), 4.03 (s, 3H), 3.81 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 152.3, 150.3 (dd, 1C, J = 3.0 Hz, 247.9 Hz), 149.7 (dd, 1C, J = 3.0 Hz, 246.4 Hz), 147.3, 134.3 (d, 1C, J = 7.7 Hz), 134.2, 124.1 (dd, 1C, J = 3.4 Hz, 6.9 Hz), 124.0, 117.9, 117.2 (dd, 2C, J = 17.6 Hz, 39.7 Hz), 116.6, 62.4, 56.8, 48.4, 45.3. APCI-MS m/z: 320.1 (MH⁺).

Example 199 2-(3,5-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23ff)

[0270] Yield 61%. ¹H NMR (CDCl₃): δ 6.99 (d, IH, J = 8.0 Hz), 6.94 (d, IH, J = 8.0 Hz), 6.74-6.69 (m, 2H), 6.59-6.54 (m, IH), 4.60 (s, 2H), 4.10 (s, 2H), 3.99 (s, 3H), 3.77 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 163.1 (dd, 2C, 7 = 12.2 Hz, 247.9 Hz), 152.3, 147.2, 141.2 (t, 1C, / = 9.1 Hz), 134.2, 124.2, 117.9, 116.7, 110.6 (dd, 2C, J = 6.5 Hz, 25.2 Hz), 102.9 (t, 1C, J = 25.2 Hz), 62.3 (d, 1C, J = 1.5 Hz), 56.6 (d, 1C, J = 1.5 Hz), 48.6, 45.5. APCI-MS m/z: 320.1 (MH⁺).

Example 200 2-(2,6-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23ggh)

[0271] Yield 79%. ¹H NMR (CDCl₃): δ 7.23-7.15 (m, 2H), 6.98 (d, IH, J = 8.4 Hz), 6.93 (d, IH, J = 8.4 Hz), 6.82 (dd, 2H, J = 7.2 Hz, 8.0 Hz), 4.78 (s, 2H), 4.11 (s, 2H), 4.01 (s, 3H), 3.78 (s, 3H). ¹³C NMR (CDCl₃): δ 166.0, 161.7 (dd, 2C, J = 7.6 Hz, 248.7 Hz), 152.2, 147.2, 134.4, 129.9 (t, 1C, 7 = 10.3 Hz), 124.5, 117.7, 116.4, 112.4, 111.4 (dd, 2C, 7 = 6.1 Hz, 19.1 Hz), 62.5, 56.7, 48.4, 33.6 (d, 1C, 7 = 3.8 Hz). APCI-MS m/z: 320.1 (MH⁺).

Example 201 2-(2-chloro-3,6-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23hh)

[0272] Yield 57%. ¹H NMR (CDCl₃): δ 7.09-7.03 (m, IH), 7.00 (d, IH, 7 = 8.0 Hz), 7.01- 6.97 (m, IH), 6.94 (d, IH, 7 = 8.0 Hz), 4.90 (s, 2H), 4.07 (s, 2H), 4.03 (d, 3H, 7 = 2.0 Hz, 2.8 Hz), 3.81 (d, 3H, 7 = 2.0 Hz). ¹³C NMR (CDCl₃): δ 166.0, 157.6 (d, 1C, 7 = 254.1 Hz), 154.8 (dd, 1C, J = 6.1 Hz, 241.1 Hz), 152.3, 147.3, 134.2, 124.3, 123.9 (d, 1C, 7 = 19.1 Hz), 117.7, 116.6, 116.4 (dd, 1C, 7 = 9.1 Hz, 23.6 Hz), 114.5 (dd, 2C, 7 = 7.6 Hz, 25.1 Hz), 62.4 (t, 1C, 7 =6.1 Hz), 56.7 (d, 1C, 7 = 7.6 Hz), 48.2 (t, 1C, 7 = 5.3 Hz), 37.2. APCI-MS m/z: 354.0 (MH⁺).

Example 202 2-(5-chloro-2,4-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one (23ii)

[0273] Yield 56%. ¹H NMR (CDCl₃): δ 7.36 (t, IH, 7 = 8.0 Hz), 7.00 (d, IH, 7 = 8.0 Hz), 6.96 (d, IH, 7 = 8.0 Hz), 6.82 (t, IH, 7 = 9.2 Hz), 4.63 (s, 2H), 4.17 (s, 2H), 3.99 (s, 3H), 3.79 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 159.0 (dd, 1C, J = 10.7 Hz, 248.0 Hz), 157.4 (dd, 1C, 7 = 12.2 Hz, 250.2 Hz), 152.3, 147.2, 134.2, 131.7 (t, 1C, 7 = 5.0 Hz), 124.2, 121.4 (dd, 1C, 7 = 3.9 Hz, 16.8 Hz), 117.9, 116.9 (d, 1C, 7 = 4.5 Hz), 116.7, 104.9 (dt, 1C, 7 = 2.3 Hz, 25.1 Hz), 62.3 (dd, 1C, 7 = 5.8 Hz, 10.7 Hz), 56.6 (dd, 1C, 7 = 6.8 Hz, 13.8 Hz), 48.8, 38.9 (d, 1C, 7 = 3.8 Hz). APCI-MS m/z: 354.0 (MH⁺).

Example 203 6,7-dimethoxy-2-(2,3,6-trifluorobenzyl)isoindolin-l-one (23jj)

[0274] Yield 52%. ¹H NMR (CDCl₃): δ 7.09-7.01 (m, IH), 7.01 (d, IH, J = 8.4 Hz), 6.96 (d, IH, J = 8.4 Hz), 6.83-6.77 (m, IH), 4.81 (s, 2H), 4.17 (s, 2H), 4.02 (s, 3H), 3.81 (s, 3H). APCI-MS m/z: 338.1 (MH⁺).

Example 204 6,7-dimethoxy-2-(2,3,4-trifluorobeπzyl)isoindolin-l-one (23kk)

[0275] Yield 54%. ¹H NMR (CDCl₃): δ 7.11-7.05 (m, IH), 7.03 (dd, IH, J = 0.8 Hz, 8.0 Hz), 6.99 (d, IH, 7 = 8.0 Hz), 6.89-6.83 (m, IH), 4.71 (s, 2H), 4.20 (s, 2H), 4.02 (d, 3H, J = 1.2 Hz), 3.82 (d, 3H, J = 1.2 Hz). APCI-MS m/z: 338.0 (MH⁺).

Example 205 6,7-dimethoxy-2-(2,4,5-trifluorobenzyl)isoindolin-l-one (2311)

[0276] Yield 49%. ^λH NMR (CDCl₃): δ 7.20-7.13 (m, IH), 7.02 (d, IH, J = 8.0 Hz), 6.98 (d, IH, J = 8.0 Hz), 6.89-6.83 (m, IH), 4.65 (s, 2H), 4.19 (s, 2H), 4.01 (d, 3H, J = 1.2 Hz), 3.81 (d, 3H, / = 1.2 Hz).

Example 206 6,7-dimethoxy-2-(3,4,5-trifluorobenzyl)isoindolin-l-one (23mm)

[0277] Yield 53%. ¹H NMR (CDCl₃): δ 7.03 (d, IH, J = 8.0 Hz), 6.98 (d, IH, J = 8.0 Hz), 6.87 (t, 2H, J = 6.8 Hz), 4.59 (s, 2H), 4.14 (s, 2H), 4.02 (s, 3H), 3.82 (s, 3H). APCI-MS m/z: 338.1 (MH⁺).

Example 207 6,7-dimethoxy-2-(perfluorobenzyI)isoindolin-l-one (23nn)

Yield 60%. ¹H NMR (CDCl₃): δ 7.20 (d, IH, J = 8.0 Hz), 6.98 (d, IH, J = 8.0 Hz), 4.21 (s, 2H), 3.99 (s, 3H), 3.84 (s, 2H), 3.80 (s, 3H). APCI-MS m/z: 374.0 (MH⁺).

Examples 208-241 [0278] The following were prepared by demethylation of intermediates 23 using General Procedure G.

Example 208 2-(3,4-dichIorobenzyl)-6,7-dihydroxyisoindolin-l-one (24uu)

[0279] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 24.0 minutes. ¹H NMR (DMSO): δ 7.54 (d, IH, 7 = 8.4 Hz), 7.47 (d, IH, J = 1.6 Hz), 7.20 (dd, IH, J = 1.6 Hz, 8.4 Hz), 6.91 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.59 (s, 2H), 4.18 (s, 2H). ¹³C NMR (DMSO): δ 168.4, 144.9, 143.4, 139.3, 132.8, 131.6, 131.3, 130.3, 130.1, 128.4, 120.0, 118.1, 114.2, 49.2, 44.5. APCI-MS m/z: 322.0, 324.0 (M-H). HRMS calcd for C₁₅H₁₂NO₃Cl₂ [MH⁺]: 324.0189. Found: 381.0183.

Example 209 (S)-6,7-dihydroxy-2-(l,2,3,4-tetrahydronaphthalen-l-yI)isoindolin-l-one (24vv)

[0280] Linear gradient of 40% B to 50% B over 30 minutes; retention time = 24.7 minutes. ¹H NMR (DMSO): δ 7.12-7.07 (m, 2H), 7.06-7.02 (m, 2H), 6.91 (d, IH, J = 8.0 Hz), 6.80 (d, IH, 7 = 7.6 Hz), 6.69 (d, IH, 7 = 7.6 Hz), 5.37-5.33 (m, IH), 4.18 (d, IH, 7 = 17.2 Hz), 3.75 (d, IH, 7 = 17.2 Hz), 2.78-2.66 (m, 2H), 1.95-1.85 (m, 3H), 1.82-1.74 (m, IH). ¹³C NMR

(DMSO): δ 169.0, 144.8, 143.4, 138.2, 135.6, 132.5, 129.6, 127.3, 127.1, 126.6, 120.0, 118.3, 114.3, 49.4, 45.8, 29.2, 28.5, 21.9. ESI-MS m/z: 294.1 (M-H). HRMS calcd for C₁₈Hi₈NO₃ [MH⁺]: 296.1281. Found: 296.1278.

Example 210 (R)-6,7-dihydroxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindoIin-l-one (24ww)

[0281] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 21.2 minutes. ¹H NMR (DMSO): δ 9.30 (s, IH), 8.83 (s, IH), 7.11-7.09 (m, IH), 7.07-7.03 (m, IH), 6.91 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 6.81 (d, IH, 7 = 7.2 Hz), 6.69 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 5.35 (dd, IH, 7 = 4.8 Hz, 8.8 Hz), 4.21 (d, IH, 7 = 16.8 Hz), 3.77 (d, IH, 7 = 16.8 Hz), 2.82-2.69 (m, 2H), 1.96-1.87 (m, 3H), 1.84-1.74 (m, IH). ¹³C NMR (DMSO): δ 169.0, 144.8, 143.3, 138.2, 135.6, 132.6, 129.7, 127.3, 127.1, 126.6, 120.0, 118.3, 114.3, 49.4, 45.8, 29.2, 28.5, 21.9. ESI-MS m/z: 294.1 (M-H). HRMS calcd for C₁₈Hi₈NO₃ [MH⁺]: 296.1281. Found: 296.1282. Example 211

(S)-2-(2,3-dihydro-lH-inden-l-yl)-6,7-dihydroxyisoindolin-l-one (24xx)

[0282] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 20.2 minutes. ¹H NMR (DMSO): δ 7.26 (d, IH, J = 7.6 Hz), 7.19 (t, IH, 7 = 7.2 Hz), 7.15 (t, IH, 7 = 7.6 Hz), 7.02 (d, IH, 7 = 7.2 Hz), 6.90 (d, IH, 7 = 7.6 Hz), 6.68 (d, IH, 7 = 7.6 Hz), 5.73 (t, IH, 7 = 8.0 Hz), 4.16 (d, IH, J = 17.2 Hz), 3.75 (d, IH, J = 17.2 Hz), 3.03-2.96 (m, IH), 2.89-2.81 (m, IH), 2.38-2.30 (m, IH), 2.08-1.99 (m, IH). ¹³C NMR (DMSO): δ 168.6, 144.7, 143.7, 143.3, 141.8, 132.5, 128.3, 127.1, 125.3, 124.3, 120.0, 118.4, 114.3, 55.9, 45.4, 30.5, 30.3. HRMS calcd for C₁₇Hi₅NO₃Na [MNa⁺]: 304.0950. Found: 304.0955. Example 212

(R)-2-(2,3-dihydro-lH-inden-l-yI)-6,7-dihydroxyisoindolin-l-one (24yy)

[0283] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 19.9 minutes. ¹H NMR (DMSO): δ 7.25 (d, IH, J = 7.6 Hz), 7.19 (t, IH, J = 7.6 Hz), 7.12 (t, IH, J = 7.2 Hz), 7.02 (d, IH, J = 7.2 Hz), 6.91 (d, IH, / = 8.0 Hz), 6.68 (d, IH, J = 8.0 Hz), 5.74 (t, IH, J = 7.6 Hz), 4.13 (d, IH, J = 16.8 Hz), 3.75 (d, IH, J = 16.8 Hz), 3.02-2.95 (m, IH), 2.89-2.81 (m, IH), 2.38-2.29 (m, IH), 2.07-1.98 (m, IH). ¹³C NMR (DMSO): δ 168.6, 144.8, 143.7, 143.3, 141.8, 132.5, 128.3, 127.1, 125.3, 124.3, 120.0, 118.4, 114.3, 55.9, 45.4, 30.5, 30.3. APCI-MS m/z: 282.1 (M-H). HRMS calcd for C₁₇Hi₅NO₃Na [MNa⁺]: 304.0950. Found: 304.0951.

Example 213 2-(4-chIoro-3-fluorobenzyl)-6,7-dihydroxyisoindolin-l-one (24zz)

[0284] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 21.3 minutes. ¹H NMR (DMSO): δ 7.50 (t, IH, J = 8.0 Hz), 7.26 (dd, IH, J = 2.0 Hz, 10.0 Hz), 7.07 (dd, IH, J = 1.2 Hz, 8.4 Hz), 6.91 (d, IH, 7 = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.60 (s, 2H), 4.19 (s, 2H). ¹³C NMR (DMSO): δ 168.5, 157.6 (d, 1C, J = 245.7 Hz), 144.9, 143.4, 140.0 (d, 1C, J = 6.1 Hz), 132.8, 131.2, 125.3 (d, 1C, / = 3.1 Hz), 120.0, 118.6 (d, 1C, J = 17.5 Hz), 118.1, 116.5 (d, 1C, J = 20.6 Hz), 114.2, 49.2, 44.7. APCI-MS m/z: 308.8 (MH⁺). HRMS calcd for C₁₅H₁₂NO₃FCl [MH⁺]: 308.0490. Found: 308.0488.

Example 214 2-(5-chIoro-2,4-difluorobenzyl)-6,7-dihydroxyisoindolin-l-one (24aaa)

[0285] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 23.7 minutes. ¹H NMR (DMSO): δ 7.49 (t, IH, 7 = 8.0 Hz), 7.48 (t, IH, 7 = 9.6 Hz), 6.91 (d, IH, J = 8.0 Hz), 6.71 (d, IH, J = 8.0 Hz), 4.62 (s, 2H), 4.20 (s, 2H). ¹³C NMR (DMSO): δ 168.3, 159.3 (dd, 1C, J = 10.7 Hz, 247.1 Hz), 157.0 (dd, 1C, 7 = 12.2 Hz, 245.2 Hz), 144.9, 143.4, 132.8, 131.7 (d, 1C, J = 6.4 Hz), 122.8 (dd, lC, 7 = 3.6 Hz, 17.5 Hz), 120.0, 118.0, 115.6 (dd, 1C, 7 = 3.5 Hz, 17.5 Hz), 114.2, 106.2 (dd, 1C, 7 = 7.5 Hz, 25.2 Hz), 49.3, 39.1. APCI-MS m/z: 326.0 (MH⁺). HRMS calcd for Ci₅Hi ₁NO₃F₂Cl [MH⁺]: 326.0396. Found: 326.0395.

Example 215 (S)-6,7-dihydroxy-2-(l-phenylethyl)isoiπdolin-l-one (24bbb)

Example 216 (R)-6,7-dihydroxy-2-(l-phenylethyl)isoindolin-l-one (24ccc)

Example 217 (S)-2-(l-(4-fluorophenyl)ethyl)-6,7-dihydroxyisoindolin-l-one (24ddd)

Example 218 (R)-2-(l-(4-fluorophenyl)ethyl)-6,7-dihydroxyisoindolin-l-one (24eee)

Example 219 (S)-6,7-dihydroxy-2-(l-(naphthalen-l-yI)ethyl)isoindolin-l-one (24fff)

Example 220 (R)-6,7-dihydroxy-2-(l-(naphthalen-l-yl)ethyl)isoindolin-l-one (24ggg)

Example 221 (S)-6,7-dihydroxy-2-(l-(naphthalen-2-yl)ethyI)isoindolin-l-one (24hhh)

Example 222 (R)-6,7-dihydroxy-2-(l-(naphthalen-2-yI)ethyl)isoindolin-l-one (24iii)

Example 223 2-(3-benzoylbeπzyl)-4-bromo-6,7-dihydroxyisoindolin-l-one (24jjj)

Example 224 2-(4-benzoylbenzyl)-6,7-dihydroxyisoindolin-l-one (24kkk)

Example 225 ό^-dihydroxy^-CS-CS-CtrifluoromethyO-SH-diazirin-S-yObenzyOisoindolin-l-one (24111)

Examples 226-257 [0286] The following were prepared by reacton with the appropriate amine using General Procedure I

Example 226 2-(3-chlorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26rr)

[0287] Yield 67%. ¹H NMR (CDCl₃): δ 7.48 (d, IH, 7 = 8.0 Hz), 7.35 (d, IH, J = 1.2 Hz), 7.27-7.24 (m, IH), 7.20-7.17 (m, 2H), 7.04 (d, IH, 7 = 8.0 Hz), 4.70 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 157.8, 147.3, 138.4, 134.4, 129.9, 128.6, 127.9, 126.8, 124.4, 121.7, 119.5, 115.9, 62.5 (d, 1C, J = 1.5 Hz), 56.6 (d, 1C, 7 = 2.3 Hz), 40.9. APCI-MS m/z: 332.0 (MH⁺).

Example 227 2-(3-bromobenzyI)-4,5-dimethoxyisoindoline-l,3-dione (26ss)

[0288] Yield 55%. ¹H NMR (CDCl₃): δ 7.51 (t, IH, J = 1.6 Hz), 7.48 (d, IH, J = 8.0 Hz), 7.35-7.30 (m, 2H), 7.12 (t, IH, J = 8.0 Hz), 7.05 (d, IH, J = 8.0 Hz), 4.70 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 157.8, 147.3, 138.6, 131.5, 130.9, 130.2, 127.3, 124.4, 122.6, 121.7, 119.5, 115.9, 62.5, 56.6, 40.9. APCI-MS m/z: 376.0 (MH⁺).

Example 228 2-(3-iodobenzyI)-4,5-dimethoxyisoindoIine-l,3-dione (26tt)

[0289] Yield 47%. ¹H NMR (CDCl₃): δ 7.71 (s, IH), 7.54 (dt, IH, J = 0.8 Hz, 7.6 Hz), 7.48 (dd, IH, / = 0.8 Hz, 8.0 Hz), 7.35 (d, IH, J = 8.0 Hz), 7.05 (d, IH, J = 8.0 Hz), 6.99 (t, IH, J = 8.0 Hz), 4.67 (s, 2H), 4.09 (d, 3H, J = 0.8 Hz), 3.89 (d, 3H, J = 0.4 Hz). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 157.8, 147.3, 138.7, 137.4, 136.9, 130.3, 127.9, 124.4, 121.7, 119.5, 115.8, 94.4, 62.5, 56.6, 40.8. APCI-MS m/z: 424.0 (MH⁺).

Example 229 2-(3,4-dichlorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26uu)

[0290] Yield 16%. ¹H NMR (CDCl₃): δ 7.48 (d, IH, J= 8.0 Hz), 7.32 (d, IH, J= 8.0 Hz), 7.32-7.29 (m, IH), 7.23-7.21 (m, IH), 7.06 (dd, IH, J = 1.6 Hz, 8.0 Hz), 74.68 (s, 2H), 4.09 (s, 3H), 3.90 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 157.9, 147.3, 138.7, 136.5, 132.6, 131.9, 130.6, 128.1, 127.0, 124.2, 119.6, 115.9, 62.5, 56.6, 40.4. Example 230

4,5-dimethoxy-2-(naphthalen-2-ylmethyl)isoindoline-l,3-dione (26vv)

[0291] Yield 39%. ¹H NMR (CDCl₃): δ 7.84 (s, IH), 7.76-7.70 (m, 3H), 7.51 (dd, IH, 7 = 1.6 Hz, 8.4 Hz), 7.42 (d, IH, J = 8.4 Hz), 7.38-7.35 (m, 2H), 6.92 (d, IH, J = 8.4 Hz), 4.90 (s, 2H), 4.08 (s, 3H), 3.79 (s, 3H). ¹³C NMR (CDCl₃): δ 167.3, 166.0, 157.6, 147.1, 134.0, 133.2, 132.8, 128.4, 127.9, 127.6, 127.5, 126.5, 126.1, 126.0, 124.5, 121.8, 119.4, 115.7, 62.5, 56.5, 41.7. APCI-MS m/z: 348.1 (MH⁺).

Example 231 (S)-4,5-dimethoxy-2-(l,2,3,4-tetrahydronaphthaIen-l-yl)isoindoIine-l,3-dione (26ww)

[0292] Yield 8%. ¹H NMR (CDCl₃): δ 7.50 (d, IH, 7 = 8.4 Hz), 7.10-7.07 (m, 3H), 7.04- 7.00 (m, IH), 6.92-6.90 (m, IH), 5.47 (dd, IH, J = 6.0, 16.8 Hz), 4.09 (s, 3H), 3.92 (s, 3H), 3.02-2.94 (m, IH), 2.80-2.76 (m, IH), 2.42-2.33 (m, IH), 2.11-2.03 (m, 2H), 1.86-1.75 (m, IH). ¹³C NMR (CDCl₃): δ 167.4, 166.0, 157.7, 147.2, 137.8, 134.8, 129.2, 126.8, 126.0, 125.8, 124.6, 121.9, 119.3, 115.8, 62.5, 56.6 (d, 1C, 7 = 3.8 Hz), 49.2, 29.4, 27.9, 22.4. α [MeOH, 21.6] = -81.3.

Example 232 (R)-4,5-dimethoxy-2-(l,2,3,4-tetrahydronaphthalen-l-yI)isoindoline-l,3-dione (26xx)

[0293] Yield 14%. ¹H NMR (CDCl₃): δ 7.50 (d, IH, J = 8.4 Hz), 7.10-7.07 (m, 3H), 7.04- 7.00 (m, IH), 6.92-6.90 (m, IH), 5.47 (dd, IH, / = 6.0, 16.8 Hz), 4.09 (s, 3H), 3.91 (s, 3H), 3.02-2.94 (m, IH), 2.99-2.93 (m, IH), 2.42-2.33 (m, IH), 2.10-2.01 (m, 2H), 1.86-1.75 (m, IH). ¹³C NMR (CDCl₃): δ 167.4, 166.0, 157.7, 147.2, 137.8, 134.8, 129.2, 126.8, 126.0, 125.8, 124.6, 121.9, 119.3, 115.8, 62.5, 56.6 (d, 1C, J = 3.8 Hz), 49.2 (d, 1C, J = 2.3 Hz), 29.4, 27.9, 22.4. α [MeOH, 21.6] = 78.2.

Example 233 (S)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dimethoxyisoindoIine-l,3-dione (26yy)

[0294] Yield 14%. ¹H NMR (CDCl₃): δ 7.46 (d, IH, J = 8.0 Hz), 7.24 (d, IH, J = 7.6 Hz), 7.20-7.16 (m, IH), 7.12-7.06 (m, 2H), 7.05 (d, IH, J = 8.0 Hz), 5.80 (dd, IH, J = 6.8, 8.4 Hz), 4.07 (s, 3H), 3.89 (s, 3H), 3.34-3.29 (m, IH), 2.98-2.90 (m, IH), 2.53-2.40 (m, 2H), ¹³C NMR (CDCl₃): δ 167.3, 165.9, 157.7, 147.1, 143.8, 140.6, 127.9, 126.4, 124.8, 124.6, 123.3, 121.9, 119.3, 115.8, 62.5, 56.6, 54.7, 31.1, 29.5. APCI-MS m/z: 324.1 (MH⁺). α [CHCl₃, 22.8] = -127.7.

Example 234 (R)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dimethoxyisoindoline-l,3-dione (26zz)

[0295] Yield 15%. ¹H NMR (CDCl₃): δ lΛl (d, IH, J = 8.0 Hz), 7.25-7.17 (m, 2H), 7.12- 7.06 (m, 2H), 7.06 (d, IH, J = 8.0 Hz), 5.81 (dd, IH, J = 7.2, 8.4 Hz), 4.08 (s, 3H), 3.90 (s, 3H), 3.37-3.29 (m, IH), 2.99-2.91 (m, IH), 2.50-2.42 (m, 2H), ¹³C NMR (CDCl₃): δ 167.3, 165.9, 157.7, 147.1, 143.8, 140.6, 127.9, 126.4, 124.8, 124.6, 123.3, 122.0, 119.3, 115.8, 62.5, 56.6, 54.7, 31.1, 29.5. APCI-MS m/z: 324.1 (MH⁺). α [CHCl₃, 22.5] = -119.4. Example 235 2-(2,3-dihydro-lH-inden-2-yl)-4,5-dimethoxyisoindoline-l,3-dione (26aaa)

[0296] Yield 23%. ¹H NMR (CDCl₃): δ 7.50 (d, IH, 7= 8.0 Hz), 7.19-7.13 (m, 4H), 7.07 (d, IH, 7 = 8.0 Hz), 5.11-5.06 (m, IH), 4.10 (s, 3H), 3.92 (s, 3H), 3.59 (dd, 2H, J = 9.2 Hz, 15.2 Hz), 3.12 (dd, 2H, J = 8.8 Hz, 15.2 Hz). ¹³C NMR (CDCl₃): δ 167.7, 166.3, 157.7, 147.1, 140.9, 124.8, 126.6 (2C), 134.6, 124.3 (2C), 121.9, 119.3, 115.8, 62.5 (d, 1C, J = 3.1 Hz), 56.6 (d, 1C, 7 = 4.6 Hz), 49.9, 36.0 (2C). APCI-MS m/z: 324.0 (MH⁺).

Example 236 2-(2-fluorobenzyI)-4,5-dimethoxyisoindoline-l,3-dione (26bbb)

[0297] Yield 61%. ¹H NMR (CDCl₃): δ 7.49 (d, IH, 7= 8.0 Hz), 7.28 (dt, IH, J= 1.2 Hz, 7.6 Hz), 7.21-7.15 (m, IH), 7.06 (d, IH, 7 = 8.0 Hz), 7.03-6.96 (m, 2H), 4.83 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 160.5 (d, 1C, 7 = 247.2 Hz), 157.7, 147.3, 130.0 (d, 1C, 7 = 3.8 Hz), 129.4 (d, 1C, 7 = 8.3 Hz), 124.4, 124.1 (d, 1C, 7 = 3.0 Hz), 123.3 (d, 1C, 7 = 14.5 Hz), 121.7, 119.5, 115.8, 115.4 (d, 1C, 7 = 21.3 Hz), 62.5, 56.6, 35.2 (d, 1C, 7 = 4.6 Hz). APCI-MS m/z: 316.1 (MH⁺).

Example 237 2-(4-chloro-3-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26ccc)

[0298] Yield 56%. ¹H NMR (CDCl₃): δ 7.50 (d, IH, 7 = 8.0 Hz), 7.28 (t, IH, 7 = 8.0 Hz), 7.18 (dd, IH, 7 = 2.0 Hz, 9.6 Hz), 7.13-7.10 (m, IH), 7.07 (d, IH, 7 = 8.0 Hz), 4.70 (s, 2H), 4.10 (s, 3H), 3.91 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 157.9 (d, 1C, 7 = 248.7 Hz), 157.8, 147.4, 137.2 (d, 1C, 7 = 6.1 Hz), 130.7 (d, 1C, 7 = 3.1 Hz), 125.0, 124.3, 121.7, 120.4 (d, 1C, 7 = 17.5 Hz), 119.6 (d, 1C, 7 = 6.9 Hz), 116.9 (dd, 1C, 7 = 5.7 Hz, 21.4 Hz), 115.9, 62.5 (d, 1C, 7 = 6.9 Hz), 56.6 (d, 1C, 7 = 9.2 Hz), 40.6 (t, 1C, 7 = 8.4 Hz). APCI-MS m/z: 350.0 (M⁺).

Example 238 2-(2-chIoro-4-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26ddd)

[0299] Yield 29%. ¹H NMR (CDCl₃): δ 7.50 (d, IH, 7 = 8.0 Hz), 7.20 (dd, IH, 7 = 6.0 Hz, 8.8 Hz), 7.08 (s, IH), 7.06 (dd, IH, 7 = 2.4 Hz, 8.0 Hz), 6.86 (dt, IH, 7 = 2.8 Hz, 8.8 Hz), 4.84 (s, 2H), 4.09 (s, 3H), 3.90 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 161.7 (d, 1C, J = 247.7 Hz), 157.9, 147.3, 133.8 (d, 1C, 7= 9.9 Hz), 130.3 (d, 1C, 7 = 9. I Hz), 129.6 (d, 1C, 7 = 3.1 Hz), 124.3, 121.6, 119.6, 116.9 (d, 1C, 7 = 25.2 Hz), 115.9, 114.1 (d, 1C, 7 = 21.4 Hz), 62.5 (d, 1C, 7 = 4.6 Hz), 56.6 (d, 1C, 7= 6.1 Hz), 38.7 (t, 1C, 7 = 7.2 Hz). APCI-MS m/z: 350.0 (MH⁺).

Example 239 2-(5-chIoro-2-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26eee)

[0300] Yield 59%. ¹H NMR (CDCl₃): δ 7.51 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 7.23 (dd, IH, 7 = 2.8 Hz, 10.8 Hz), 7.17-7.13 (m, IH), 7.07 (d, IH, 7 = 8.0 Hz), 6.94 (d, IH, 7 = 9.2 Hz), 4.79 (s, 2H), 4.10 (d, 3H, 7 = 0.8 Hz), 3.90 (s, 3H). ¹³C NMR (CDCl₃): δ 166.9, 165.7, 159.0 (d, 1C, 7 = 246.4 Hz), 157.9, 147.4, 129.7 (d, 1C, 7 = 3.8 Hz), 129.3 (d, 1C, 7 = 8.4 Hz), 129.2 (d, 1C, 7 = 3.9 Hz), 125.1 (d, 1C, 7= 16.8 Hz), 124.3, 121.6, 119.6, 116.8 (d, 1C, 7 = 12.9 Hz), 115.9, 62.5, 56.6, 34.8. APCI-MS m/z: 350.0 (MH⁺). Example 240 2-(2-chloro-6-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26fff)

151D-143 [0301] Yield 41%. ¹H NMR (CDCl₃): δ 7.49 (d, IH, 7 = 8.0 Hz), 7.20-7.12 (m, 2H), 7.06 (d, IH, / = 8.0 Hz), 6.97-6.93 (m, IH), 4.94 (d, 2H, 7 = 1.2 Hz), 4.08 (s, 3H), 3.90 (s, 3H). ¹³C NMR (CDCl₃): δ 166.7, 165.5, 161.9 (d, 1C, 7= 250.2 Hz), 157.7, 147.2, 135.3 (d, 1C, 7 = 5.3 Hz), 129.7 (d, 1C, J = 9.9 Hz), 125.4, 124.5, 121.7, 121.4 (d, 1C, J = 16.0 Hz), 119.4 (dd, 1C, 7 =1.6 Hz, 9.2 Hz), 115.8 (d, 1C, 7 = 4.6 Hz), 114.2 (d, 1C, J = 22.9 Hz), 62.5 (d, 1C, 7 = 7.7 Hz), 56.6 (dd, 1C, 7 = 9.6 Hz, 21.3 Hz), 33.8 (dd, 1C, 7 = 13.0 Hz, 16.8 Hz). APCI-MS m/z: 349.9 (MH⁺).

Example 241 2-(3-chloro-2-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26ggg)

[0302] Yield 55%. ¹H NMR (CDCl₃): δ 7.51 (dd, IH, 7 = 0.8 Hz, 8.0 Hz), 7.28-7.23 (m, IH), 7.21-7.18 (m, IH), 7.07 (d, IH, 7 = 8.0 Hz), 6.97 (t, IH, 7 = 8.0 Hz), 4.85 (s, 2H), 4.10 (d, 3H, 7 = 0.8 Hz), 3.91 (s, 3H). ¹³C NMR (CDCl₃): δ 166.9, 165.7, 157.8, 156.0 (d, 1C, 7 = 248.7 Hz), 147.3, 130.0 (d, 1C, 7 = 4.6 Hz), 128.3, 125.0 (d, 1C, 7 = 14.5 Hz), 124.5 (d, 1C, 7 = 4.5 Hz), 124.3, 121.6, 121.2 (d, 1C, 7 = 17.5 Hz), 119.5 (dd, 1C, 7 = 1.6 Hz, 9.2 Hz), 115.9 (d, 1C, 7 = 3.0 Hz), 62.6 (d, 1C, 7 = 7.6 Hz), 56.5 (d, 1C, 7 = 10.7 Hz), 35.3 (dd, 1C, 7 = 12.2 Hz, 16.8 Hz). APCI-MS m/z: 350.0 (MH⁺).

Example 242 2-(3-fluoro-4-methylbenzyI)-4,5-dimethoxyisoindoline-l,3-dione (26hhh)

[0303] Yield 71%. ¹H NMR (CDCl₃): (57.47 (d, IH, / = 8.0 Hz), 7.04 (d, IH, / = 8.0 Hz), 7.04-7.00 (m, 2H), 4.68 (s, 2H), 4.08 (s, 3H), 3.88 (s, 3H), 2.16 (d, 3H, / = 1.60 Hz). ¹³C NMR (CDCl₃): δ 167.2, 165.9, 161.1 (d, 1C, / = 244.2 Hz), 157.7, 147.2, 136.0 (d, IC, / = 6.9 Hz), 131.5 (d, 1C, / = 16.8 Hz), 124.4, 124.1 (dd, 1C, / = 22.1 Hz, 36.7 Hz), 121.8, 119.6, 119.3, 115.8 (dd, 1C, /= 2.3 Hz, 18.6 Hz), 115.1 (dd, IC, / = 19.0 Hz, 24.4 Hz), 62.5 (d, 1C, / = 21.3 Hz), 56.5 (d, 1C, / = 22.1 Hz), 40.8 (t, 1C, 7 = 19.8 Hz), 14.2 (dd, 1C, / = 3.8 Hz, 16.8 Hz). APCI-MS m/z: 330.1 (MH⁺).

Example 243 2-(4-fluoro-3-methylbenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26iii)

[0304] Yield 77 %. ¹H NMR (CDCl₃): δ 7.48 (d, IH, / = 8.0 Hz), 7.21-7.16 (m, 2H), 7.04 (d, IH, J = 8.0 Hz), 6.87 (t, IH, / = 8.0 Hz), 4.67 (s, 2H), 4.09 (s, 3H), 3.89 (s, 3H), 2.18 (d, 3H, / = 2.0 Hz). ¹³C NMR (CDCl₃): δ 167.3, 165.9, 160.8 (d, 1C, / = 243.3 Hz), 157.7, 147.2, 132.0 (d, 1C, / = 3.8 Hz), 131.9 (t, 1C, / = 5.8 Hz), 127.8, 125.0 (d, 1C, / = 17.5 Hz), 124.5, 121.9, 119.4 (d, 1C, / = 8.4 Hz), 115.7, 115.0 (d, 1C, / = 22.1 Hz), 62.5 (d, 1C, / = 6.8 Hz), 56.5 (d, 1C, / = 9.2 Hz), 40.8 (t, 1C, / = 7.6 Hz), 14.5. APCI-MS m/z: 330.1 (MH⁺).

Example 244 2-(3-bromo-4-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26jjj)

[0305] Yield 67%. ¹H NMR (CDCl₃): δ 7.55 (dd, IH, / = 2.4 Hz, 6.4 Hz), 7.47 (d, IH, / = 8.0 Hz), 7.33-7.29 (m, IH), 7.05 (d, IH, / = 8.0 Hz), 6.98 (t, IH, / = 8.4 Hz), 4.67 (s, 2H), 4.08 (s, 3H), 3.89 (s, 3H). ¹³C NMR (CDCl₃): δ 167.1, 165.8, 158.6 (d, 1C, 7 = 246.4 Hz), 157.8, 147.3, 133.9 (d, 1C, 7= 3.9 Hz), 133.8, 129.5 (d, 1C, J = 7.6 Hz), 124.3, 121.7, 119.6, 116.5 (d, 1C, J = 22.9 Hz), 115.9, 109.0 (d, 1C, 7 = 10.6 Hz), 62.5, 56.6, 40.3. APCI-MS m/z: 393.9 (MH⁺).

Example 245 2-(2,3-difluorobenzyl)-4,5-dimethoxyisoindoIine-l,3-dione (26kkk)

[0306] Yield 48%. ¹H NMR (CDCl₃): δ 7.52 (d, IH, 7 = 8.0 Hz), 7.08 (d, IH, 7 = 8.0 Hz), 7.06-7.02 (m, 2H), 7.00-6.94 (m, IH), 4.86 (s, 2H), 4.11 (s, 3H), 3.92 (s, 3H). ¹³C NMR (CDCl₃): δ 167.0, 165.7, 157.8, 150.5 (dd, 1C, J = 12.2 Hz, 247.1 Hz), 148.7 (dd, 1C, 7 =

11.9 Hz, 249.7 Hz), 147.4, 125.7 (d, 1C, 7 = 12.4 Hz), 124.6 (t, 1C, J = 3.0 Hz), 124.3, 124.0 (dd, 1C, 7 = 4.6 Hz, 6.8 Hz), 121.6, 119.6, 116.6 (d, 1C, 7 = 17.5 Hz), 115.9, 62.6, 56.6, 34.8. APCI-MS m/z: 334.1 (MH⁺).

Example 246 2-(2,4-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26111)

[0307] Yield 51%. ¹H NMR (CDCl₃): δ 7.48 (d, IH, J = 8.0 Hz), 7.33-7.28 (m, IH), 7.06 (d, IH, 7 = 8.0 Hz), 6.78-6.70 (m, 2H), 4.78 (s, 2H), 4.08 (s, 3H), 3.89 (s, 3H). ¹³C NMR (CDCl₃): δ 167.0, 165.7, 162.5 (dd, 1C, 7 = 11.4 Hz, 247.2 Hz), 160.6 (dd, 1C, 7 = 12.3 Hz, 250.3 Hz), 157.8, 147.3, 131.3 (dd, 1C, 7 = 5.4 Hz, 10.0 Hz), 124.3, 121.7, 119.5, 119.3 (dd, 1C, 7= 3.8 Hz, 14.5 Hz), 115.9, 111.3 (dd, 1C, 7 = 3.8 Hz, 21.3 Hz), 103.9 (t, 1C, 7 = 24.4 Hz), 62.5 (d, 1C, 7 = 2.3 Hz), 56.6 (d, 1C, 7 = 3.1 Hz), 34.6 (d, 1C, J = 3.9 Hz). APCI-MS m/z: 334.1 (MH⁺).

Example 247 2-(2,5-difluorobenzyl)-4,S-dimethoxyisoindoline-l,3-dione (26mmm)

[0308] Yield 37%. ¹H NMR (CDCl₃): δ 7.48 (d, IH, J = 8.0 Hz), 7.06 (d, IH, J = 8.0 Hz), 6.97-6.90 (m, 2H), 6.87-6.81 (m, IH), 4.78 (s, 2H), 4.07 (s, 3H), 3.88 (s, 3H). ¹³C NMR (CDCl₃): δ 166.9, 165.7, 158.5 (dd, 1C, 7 = 3.0 Hz, 241.1 Hz), 157.8, 156.4 (dd, 1C, J = 3.0 Hz, 241.8 Hz), 147.3, 124.9 (dd, 1C, J = 7.6 Hz, 17.5 Hz), 124.2, 121.6, 119.6, 116.5 (d, 1C, J = 8.4 Hz, 23.7 Hz), 116.2 (dd, 1C, J = 4.6 Hz, 24.4 Hz), 115.9, 115.7 (dd, 1C, J = 8.4 Hz, 24.4 Hz), 62.5 (d, 1C, J = 2.3 Hz), 56.6 (d, 1C, J = 3.8 Hz), 34.9 (d, 1C, J = 3.8 Hz). APCI- MS m/z: 334.1 (MH⁺).

Example 248 2-(3,4-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26nnn)

[0309] Yield 48%. ¹H NMR (CDCl₃): δ 7.49 (d, IH, J = 8.0 Hz), 7.24-7.19 (m, IH), 7.13- 7.10 (m, IH), 7.06 (d, IH, J = 8.0 Hz), 7.04-7.00 (m, IH), 4.69 (s, 2H), 4.09 (s, 3H), 3.90 (s, 3H). APCI-MS m/z: 334.1 (MH⁺).

Example 249 2-(3,5-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26ooo)

[0310] Yield 54%. ¹H NMR (CDCl₃): δ 7.47 (d, IH, J = 8.0 Hz), 7.05 (d, IH, J = 8.0 Hz), 6.88-6.83 (m, 2H), 6.64-6.59 (m, IH), 4.68 (s, 2H), 4.07 (s, 3H), 3.88 (s, 3H). ¹³C NMR (CDCl₃): δ 167.0, 165.7, 163.0 (dd, 2C, J = 13.0 Hz, 248.0 Hz), 157.9, 147.3, 140.1 (t, 1C, J = 9.1 Hz), 124.2, 121.6, 119.6 (d, 1C, 7 = 1.6 Hz), 115.9, 111.3 (dd, 2C, 7 = 6.8 Hz, 18.3 Hz), 103.2 (dd, 1C, J = 4.5 Hz, 25.2 Hz), 62.5 (d, 1C, J = 4.6 Hz), 56.5 (d, 1C, / = 5.3 Hz), 40.6. APCI-MS m/z: 334.1 (MH⁺).

Example 250 2-(2,6-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26ppp)

[0311] Yield 77%. ¹H NMR (CDCl₃): δ 7.45 (dd, IH, J = 3.2 Hz, 8.0 Hz), 7.19-7.12 (m, IH), 7.04 (d, IH, 7 = 8.0 Hz), 6.82-6.76 (m, 2H), 4.83 (s, 2H), 4.05 (s, 3H), 3.87 (s, 3H). ¹³C NMR (CDCl₃): δ 166.6, 165.4, 161.5 (dd, 2C, J = 7.7 Hz, 248.7 Hz), 157.7, 147.2, 129.6 (t, 1C, J = 10.7 Hz), 124.4, 121.7, 119.4, 115.8, 111.8 (t, 1C, J = 17.6 Hz), 111.3 (dd, 2C, J = 6.1 Hz, 19.1 Hz), 62.5, 56.5, 30.0 (t, 1C, J = 4.6 Hz). APCI-MS m/z: 334.1 (MH⁺).

Example 251 2-(2-chloro-3,6-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26qqq)

[0312] Yield 56%. ¹H NMR (CDCl₃): δ 7.47 (d, IH, / = 8.0 Hz), 7.06 (d, IH, / = 8.0 Hz), 7.04-7.00 (m, IH), 6.95-6.89 (m, IH), 4.92 (s, 2H), 4.06 (s, 3H), 3.89 (s, 3H). ¹³C NMR

(CDCl₃): δ 166.6, 165.4, 157.5 (dd, 1C, J= 2.3 Hz, 248.7 Hz), 157.8, 154.7 (dd, 1C, J = 3.0 Hz, 243.4 Hz), 147.2, 124.3, 123.0 (d, 1C, J = 17.5 Hz), 121.6, 119.5 (d, 1C, / = 6.9 Hz), 116.1 (dd, 1C, 7 = 9.9 Hz, 23.1 Hz), 115.9 (2C), 114.4 (dd, 1C, / = 7.6 Hz, 25.2 Hz), 62.5 (d, 1C, / = 6.8 Hz), 56.6 (d, 1C, J = 9.1 Hz), 33.7. APCI-MS m/z: 368.0 (MH⁺). Example 252

2-(5-chIoro-2,4-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione (26rrr)

[0313] Yield 68%. ¹H NMR (CDCl₃): δ 7.46 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.33 (t, IH, / = 7.8 Hz), 7.05 (d, IH, J = 8.0 Hz), 6.82 (t, IH, J = 9.0 Hz), 4.72 (s, 2H), 4.06 (s, 3H), 3.88 (s, 3H). ¹³C NMR (CDCl₃): δ 166.8, 165.6, 158.8 (dd, 1C, J = 10.3 Hz, 249.4 Hz), 157.9, 157.4 (dd, 1C, J = 12.2 Hz, 250.2 Hz), 147.3, 131.4 (dd, 1C, J = 5.0 Hz, 10.3 Hz), 124.1, 121.5, 120.7 (dd, 1C, / = 4.5 Hz, 16.0 Hz), 119.6 (d, 1C, J = 10.6 Hz), 116.4 (dd, 1C, J = 4.2 Hz, 11.1 Hz), 116.0 (d, 1C, J = 4.5 Hz), 105.1 (dt, 1C, J = 3.0 Hz, 25.2 Hz), 62.4 (dd, 1C, J = 9.9 Hz, 19.1 Hz), 56.6 (dd, 1C, J = 12.2 Hz, 24.4 Hz), 34.3 (dd, 1C, 7 = 10.6 Hz, 14.4 Hz). APCI-MS m/z: 368.0 (MH⁺).

Example 253 4,5-dimethoxy-2-(2,3,6-trifluorobenzyl)isoindoline-l,3-dione (26sss)

[0314] Yield 63%. ¹H NMR (CDCl₃): δ 7.46 (d, IH, J = 8.0 Hz), 7.05 (d, IH, J = 8.0 Hz), 7.05-6.97 (m, IH), 6.78-6.72 (m, IH), 4.84 (s, 2H), 4.06 (s, 3H), 3.88 (s, 3H). APCI-MS m/z: 352.1 (MH⁺). Example 254

4,5-dimethoxy-2-(2,3,4-trifluorobenzyl)isoindoline-l,3-dione (26ttt)

[0315] Yield 73%. ¹H NMR (CDCl₃): δ 7.46 (d, IH, J = 8.0 Hz), 7.06 (d, IH, J = 8.0 Hz), 7.03-7.02 (m, IH), 6.86-6.80 (m, IH), 4.77 (s, 2H), 4.06 (s, 3H), 389 (s, 3H). Example 255

4,5-dimethoxy-2-(2,4,5-trifluorobenzyl)isoindoline-l,3-dione (26uuu)

[0316] Yield 24%. ¹H NMR (CDCl₃): δ 7.51 (dd, IH, 7 = 0.8 Hz, 8.4 Hz), 7.19-7.12 (m, IH), 7.08 (d, IH, J = 8.4 Hz), 6.90-6.84 (m, IH), 4.76 (s, 2H), 4.10 (s, 3H), 3.91 (s, 3H). APCI-MS m/z: 352.1 (MH⁺). Example 256

4,5-dimethoxy-2-(3,4,5-trifluorobenzyl)isoindoline-l,3-dione (26vvv)

[0317] Yield 64%. ¹H NMR (CDCl₃): δ 7.51 (d, IH, J = 8.0 Hz), 7.08 (d, IH, J = 8.0 Hz), 7.02 (t, 2H, J= 6.8 Hz), 4.67 (s, 2H), 4.10 (s, 3H), 3.92 (s, 3H). APCI-MS m/z: 352.1 (MH⁺). Example 257

4,5-dimethoxy-2-(perfluorobenzyI)isoindoline-l,3-dione (26www)

[0318] Yield 14%. ¹H NMR (DMSO): δ 7.49 (d, IH, / = 8.4 Hz), 7.08 (d, IH, J = 8.4 Hz), 4.86 (d, 2H, J= 0.8 Hz), 4.08 (s, 3H), 3.91 (s, 3H). APCI-MS m/z: 388.0 (MH⁺). Examples 258-265

[0319] The following were prepared by demethylation of intermediates 26 using General Procedure G.

Example 258 2-(3,4-dichlorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27rr)

[0320] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 27.5 minutes. ¹H NMR (DMSO): 7.53 (d, IH, J = 8.4 Hz), 7.49 (d, IH, J = 2.0 Hz), 7.20 (dd, IH, J = 2.0, 8.4 Hz), 7.14 (d, IH, J = 1.6 Hz), 7.01 (d, IH, J = 1.6 Hz), 4.62 (s, 2H). ¹³C NMR (CDCl₃): δ 167.6, 166.7, 153.0, 144.8, 138.7, 131.5, 131.2, 130.4, 129.9, 128.1, 122.6, 119.2, 116.3, 116.1, 39.8. ESI-MS m/z: 336.0 (M-H). HRMS calcd for Ci₅H₈NO₄Cl₂ [M-H]: 335.9836. Found: 335.9845.

Example 259 (S)-4,5-dihydroxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindoline-l,3-dione (27ss)

[0321] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 23.5 minutes. ¹H NMR (CD₃OD): δ lΛl (d, IH, J = 8.0 Hz), 7.07-7.06 (m, 2H), 7.01 (d, IH, J = 8.0 Hz), 7.01-6.98 (m, 2H), 6.85 (d, IH, J = 7.6 Hz), 5.37 (dd, IH, J = 6.4, 10.8 Hz), 2.96-2.88 (m, IH), 2.82-2.76 (m, IH), 2.41-2.32 (m, IH), 2.10-2.00 (m, 2H), 1.85-1.76 (m, IH). ¹³C NMR (CD₃OD): δ 168.2, 167.7, 152.3, 137.5, 135.0, 128.7, 126.3, 125.7, 125.2, 122.4, 121.8, 118.6, 115.8, 115.5, 48.8, 29.0, 27.6, 22.3. ESI-MS m/z: 308.1 (M-H). HRMS calcd for Ci₈H₁₄NO₄ [M-H]: 308.0928. Found: 308.0928.

Example 260 (R)-4,5-dihydroxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindoline-l,3-dione (27tt)

[0322] Linear gradient of 40% B to 60% B over 30 minutes; retention time = 22.1 minutes. ¹U NMR (DMSO): δ 10.81 (s, IH), 10.08 (s, IH), 7.11 (d, IH, J = 7.6 Hz), 7.07 (d, IH, J = 4.0 Hz), 7.01 (d, IH, / = 7.6 Hz), 7.01-6.97 (m, IH), 6.80 (d, IH, J = 8.0 Hz), 5.23 (dd, IH, J = 6.4, 10.8 Hz), 2.84-2.70 (m, 2H), 2.27-2.19 (m, IH), 2.02-1.86 (m, 2H), 1.74-1.70 (m, IH). ESI-MS m/z: 308.1 (M-H). HRMS calcd for C₁₈H₁₄NO₄ [M-H]: 308.0928. Found: 308.0921.

Example 261 (S)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dihydroxyisoindoline-l,3-dione (27uu)

[0323] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 20.8 minutes. ¹H NMR (DMSO): δ 7.21 (d, IH, J = 7.6 Hz), 7.15 (t, IH, J = 7.2 Hz), 7.11-7.04 (m, 2H), 7.02-6.99 (m, 2H), 5.61 (t, IH, J = 8.0 Hz), 3.14-3.07 (m, IH), 2.90-2.82 (m, IH), 2.38-2.28 (m, 2H). ¹³C NMR (DMSO): δ 167.5, 166.7, 152.8, 144.5, 143.7, 141.6, 127.9, 126.8, 125.0, 123.4, 122.6, 119.2, 116.1, 116.0, 54.0, 30.9, 29.6. ESI-MS m/z: 294.1 (M-H). HRMS calcd for C₁₇H₁₂NO₄ [M-H]: 294.0766. Found: 294.0767.

Example 262 (R)-2-(2,3-dihydro-lH-inden-l-yI)-4,5-dihydroxyisoindoline-l,3-dione (27vv)

[0324] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 21.3 minutes. ¹H NMR (DMSO): δ 10.8 (brs, IH), 10.0 (brs, IH), 7.22 (d, IH, J = 7.2 Hz), 7.15 (t, IH, J = 7.2 Hz), 7.11-7.05 (m, 2H), 7.00 (dd, 2H, J = 1.2 Hz, 8.0 Hz), 5.60 (t, IH, / = 8.0 Hz), 3.15- 3.07 (m, IH), 2.91-2.83 (m, IH), 2.40-2.26 (m, 2H). ¹³C NMR (DMSO): δ 167.5, 166.7, 152.8, 144.5, 143.7, 141.6, 127.9, 126.8, 125.0, 123.4, 122.6, 119.2, 116.1, 116.0, 54.0, 30.9, 29.6. HRMS calcd for C₁₇H₁₂NO₄ [M-H]: 294.0766. Found: 294.0762.

Example 263 2-(4-chloro-3-fluorobenzyl)-4,5-dihydroxyisoindoline- 1 ,3-dione (27ww)

[0325] Linear gradient of 40% B to 55% B over 30 minutes; retention time = 24.0 minutes. ¹H NMR (DMSO): δ 7.47 (t, IH, 7 = 8.0 Hz), 7.27 (dd, IH, J = 1.6 Hz, 10.0 Hz), 7.14 (d, IH, /= 8.0 Hz), 7.07 (dd, IH, J = 1.6 Hz, 8.0 Hz), 7.02 (d, IH, J = 8.0 Hz), 4.63 (s, 2H). ¹³C NMR (DMSO): δ 167.6, 166.7, 157.5 (d, 1C, 7 = 245.6 Hz), 152.9, 147.7, 139.3 (d, 1C, J = 6.8 Hz), 131.1, 124.9 (d, 1C, 7 = 3.9 Hz), 122.6, 119.2, 118.7 (d, 1C, J = 17.5 Hz), 116.3, 116.2 (d, 2C, J = 14.5 Hz), 39.7. MALDI-MS m/z: 321.62 (M⁺). HRMS calcd for C₁₅H₈NO₄FCl [M-H]: 320.0126. Found: 320.0129.

Example 264 2-(5-chloro-2,4-difluorobenzyI)-4,5-dihydroxyisoindoline-l,3-dione (27xx)

[0326] Linear gradient of 40% B to 70% B over 30 minutes; retention time = 20.2 minutes. ¹H NMR (DMSO): δ 7.51 (t, IH, / = 8.0 Hz), 7.47 (t, IH, J = 9.6 Hz), 7.13 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.01 (dd, IH, J = 0.8 Hz, 8.0 Hz), 4.64 (s, 2H). ¹³C NMR (DMSO): δ 161 A, 166.6, 158.8 (d, 1C, J = 248.0 Hz), 157.0 (d, 1C, J = 247.9 Hz), 152.9, 144.7, 131.4 (d, 1C, 7 = 5.3 Hz), 122.6, 122.3 (d, 1C, 7 = 16.0 Hz), 119.2, 116.2 (2C), 115.6 (d, 1C, 7 = 10.5 Hz), 106.1 (t, 1C, 7 = 25.2 Hz), 34.4. MALDI-MS m/z: 339.62 (MH⁺). HRMS calcd for C₁₅H₇NO₄F₂Cl [M-H]: 338.0032. Found: 338.0035.

Example 265 2-(4-benzoyIbenzyl)-4,5-dihydroxyisoindoline-l,3-dione (27yy)

Example 266 2-(3-chloro-2-fluorobenzyl)-4,5-dihydroxyisoindoIine-l,3-dione (27zz)

[0327] Yield 55%. ¹H NMR (CDCl₃): δ 7.51 (dd, IH, J = 0.8 Hz, 8.0 Hz), 7.28-7.23 (m, IH), 7.21-7.18 (m, IH), 7.07 (d, IH, 7 = 8.0 Hz), 6.97 (t, IH, J = 8.0 Hz), 4.85 (s, 2H), 4.10 (d, 3H, / = 0.8 Hz), 3.91 (s, 3H). ¹³C NMR (CDCl₃): δ 166.9, 165.7, 157.8, 156.0 (d, 1C, J = 248.7 Hz), 147.3, 130.0 (d, 1C, J = 4.6 Hz), 128.3, 125.0 (d, 1C, J = 14.5 Hz), 124.5 (d, 1C, J = 4.5 Hz), 124.3, 121.6, 121.2 (d, 1C, J = 17.5 Hz), 119.5 (dd, 1C, J= 1.6 Hz, 9.2 Hz), 115.9 (d, 1C, J = 3.0 Hz), 62.6 (d, 1C, / = 7.6 Hz), 56.5 (d, 1C, / = 10.7 Hz), 35.3 (dd, 1C, J = 12.2 Hz, 16.8 Hz). APCI-MS m/z: 350.0 (MH⁺).

Example 267 2-(3-chloro-2-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione (41)

[0328] Phthalimide potassium salt (3.20 g, 17.3 mmol) was added to the solution of 2,3- difluorobenzylbromide (2.0 mL, 15.7 mmol) in DMF (anhydrous, 15 mL) with several portions. The mixture was stirred at 12O⁰C for 2.5 hours and cooled to room temperature. The reaction was quenched with ice and extracted with ethyl acetate (200 mL). Organic phase was washed with brine (6 x 30 mL) and dried by anhydrous sodium sulfate. The solution was filtered and concentrated and the residue was purified with silica gel column. 151D-88 (4.36 g, 92% yield) was afforded. ¹H NMR (CDCl₃): δ 7.78-7.76 (m, IH), 7.66-7.64 (m, 2H), 7.02- 6.92 (m, 3H), 4.86 (s, 2H). APCI-MS m/z: 274.1 (MH⁺).

Recombinant HIV IN and oligonucleotide substrates. [0329] Expression of the recombinant IN in Escherichia coli and subsequent purification of the protein were performed as previously reported (Leh, H. et al.. Biochemistry 2000, 39, 9285-9294; Marchand, C. et al. Methods Enzymol. 2001, 340, (Drug-Nucleic Acid Interactions), 624-633) with addition of 10% glycerol to all buffers. Preparation of oligonucleotide substrates has been described (Semenova, E. A. et al. MoI. Pharmacol. 2006, 69, 1454-1460). The preparation of the Q148C/SSS-mutant ESf is described at (Johnson et al, J. Biol. Chem. 281(l):461-467, 2006). The oligonucleotide substrates: 21top, 5'- GTGTGGAAAATCTCTAGCAGT-3'; 21bot, 5'-ACTGCTAGAGATTTTCCACAC-S' ; 19top, 5'-GTGTGGAAAATCTCTAGCA-S' ; 33dis, 5'- ATGTGGAAAATCTCTAGCAGGCTGCAGGTCGAC-3'; 16dis, 5'- CAGCAACGCAAGCTTG-3'; 30dis, 5'-GTCGACCTGCAGCCCAAGCTTGCGTTGCTG- 3', 21dis 5'-ACTGCTAG AG ATTTTCC AC AT-3' were purchased from MWG-Biotech Inc. (High Point, NC) and purified by polyacrylamide gel. The single-stranded oligonucleotides, 21top, 19top, 33dis, were 5'-end labeled with [γ³²P]-ATP (Perkin Elmer, Wellesley, MA) and T4 polynucleotide kinase (New England BioLabs, Ipswich, MA). Unincorporated nucleotide was removed using mini Quick Spin Oligo columns (Roche Diagnostics, Indianapolis, IN). IN substrates were obtained after annealing of the radiolabeled oligonucleotides with complementary non-labeled oligonucleotides: [γ³²P]-21top annealed with 21bot (21 bp substrate for measurement of 3'-P and ST reactions), [γ³²P]-19top annealed with 21bot (precleaved substrate for measurement of strand transfer reactions), and [γ³²P]-33dis annealed with 21dis, 30dis and 16 dis (disintegration substrate for measurement of disintegration).

Example 267

Recombinant HIV IN catalytic assays

[0330] 3'-P, ST and disintegration were examined using oligonucleotide-based assays (Chow et al., Science 255(5045):723-726, 1992; and Marchand et al., Methods Enzymol. 340:624-633, 2001) with modifications. Briefly, IN-DNA complexes (300 nM of IN and 20 nM of 5'-end [³²P]-labeled DNA substrate) were preformed, and the integration reactions were performed in 10 μL with 400 nM of recombinant IN, 20 nM of 5 '-end [³²P] -labeled oligonucleotide substrate and inhibitors at various concentrations. Solutions of 10% DMSO without inhibitors were used as controls. Reactions were incubated at 37 ⁰C (30 minutes) in buffer containing at a final concentration of 50 mM MOPS, pH 7.2 and 7.5 mM of divalent cations (MgCl₂ unless MnCl₂ is otherwise indicated). Reactions were stopped by addition of 20 μL of loading dye (10 mM EDTA, 98% deionized formamide, 0.025% xylene cyanol and 0.025% bromophenol blue). Reactions were heated at 95 ⁰C (1 minute) then subjected to electrophoresis in 20% polyacrylamide-7 M urea gels. Gels were dried and reaction products were visualized and quantitated with a Phosphorlmager (GE Healthcare, Little Chalfont, Buckinghamshire, UK). Densitometric analyses were performed using ImageQuant from Molecular Dynamics Inc. The concentrations at which enzyme activity was reduced by 50% (IC₅₀) were determined using "Prism" software (GraphPad Software, San Diego, CA) for nonlinear regression to fit dose-response data to logistic curve models.

IN binding to HIV DNA using the Schiff-base assay

[0331] The Schiff-base assay is performed as described (Mazumder and Pommier, Nucleic Acids Res. 23(15):2865-2871, 1995). Briefly, 300 nM recombinant IN was incubated with a test compound (at the indicated concentration) for 15 min at 37°C. Subsequently, 20 nM of 5'-end labeled substrate containing the abasic oligonucleotide (Fig. 2A) was added for 10 min at room temperature in reaction buffer described above for IN catalytic assays. A freshly prepared solution of sodium borohydride (0.1 M final concentration) was added for 5 min. An equal volume (10 μl) of 2X SDS-polyacrilamide gel electrophoresis buffer (Invitrogen, Carlsbad, CA) was added in each reaction. Reaction products were heated at 95⁰C for 1 min before analysis by electrophoresis in a 12-20% polyacrylamide gels (Invitrogen, CA, USA). Gels were dried and reaction products were quantitated using the same method as described above.

IN binding to HIV DNA using the disulfide-crosslinking assay

[0332] The disulfide crosslinking assay described in detail (Johnson et al. , J. Biol. Chem. 281(1 ):461-467, 2006). Briefly, 500 nM IN is incubated with a test compound in the buffer 20 mM Tris, pH 7.4, 10% glycerol and 7.5 mM of divalent cations (MgCl₂ or MnCl₂ as indicated) for 20 minutes. DNA (20 nM) containing a [5 '-³²P] -label on one strand and a thiol-modified cytosine on the other strand is added and reactions is capped with mefhylmethanethiosulfonate at 1 minute. Following capping, non-reducing gel loading buffer (100 mM Tris-HCl, pH 6.8, 4% SDS, 0.2% bromophenol blue, 20% glycerol) is added and samples are directly loaded on 16% tricine gels (Invitrogen, Carlsbad, CA). Gels are dried and reaction products are quantitated using the same way as described above. Determining inhibitor binding site in HIV IN using antibodies mapping

[0333] The effect of compounds to interfere with IN region 1-16 aa, 23-34 aa, 22-31+82- 101 aa and 276-288 aa is determined by performing complexes INwt - antibody against indicated epitope at 1:5 molar ratio for 20 min. Subsequently, the IC₅₀ for investigated compounds is measured as described above for 3'-P and ST reactions. Antibodies raised against epitopes (1-16 aa, 23-34 aa, 22-31+82-101 aa (8G4) and 276-288 aa) may be purchased from NIH AIDS Research and Reference Reagent Program (Germantown, MD) and were shown marginal inhibitory effect on IN enzymatic functions (Nilsen et ah, J. Virol. 70(3): 1580-1587, 1996; and Yi et al, J. Biol. Chem. 277(14):12164-12174, 2002).

Cell-based Assays.

Inhibition of HIV-induced cytopathic effect in cell culture

[0334] The MT-2 cells were grown in RPMI- 1640 medium with GlutaMAX™, supplemented with 10% (v/v) heat-inactivated fetal bovine serum (both from Gibco, Invitrogen corporation, Carlsbad). The cells were maintained at 37⁰C in a humidified atmosphere of 5% CO₂ in air. Every 4-5 days, cells were spun down and seeded at 2 x 10⁵ cells/ml in new cell culture flasks. HIV (HTLV-IIIB isolate) was obtained from Advanced Biotechnology Incorporated (Columbia, MD). The virus stock [3,2 x 10⁴ CdD₅₀ (50% cell culture infective dose) per ml as determinated for MT-2 cells] was stored at - 70⁰C until used. Stock solutions of compounds were diluted using medium directly into 96-well assay plate (Costar, Corning inc, Corning, NY).

[0335] MT-2 cells (5 x 10⁵ cells/ml) were infected with 100 CdD₅₀ or mock-infected. Subsequently test compounds were added at various concentrations 0.15 - 111 μM. The cell cultures were incubated at 37°C in a humidified atmosphere of 5% CO₂ in air. Four days after infection the viability of mock- and HIV-infected cells was examined spectrophotometrically by the CellTiter 96 Non-Radioactive Cell proliferation assay (Promega, Madison, WI) and also confirmed microscopically in a hemacytometer by the trypan blue exclusion method. The percent cell viability in drug treated uninfected and infected cells was determined based on the viability of the uninfected control drug treated cells. The concentration of drug required to inhibit approximately 50% of the HIV-I induced cytotoxicity was calculated from the plot of compound concentration verses the percent viable cells. [0336] Human embryonyl kidney cell culture line 293 was obtained from the American Type Culture Collection (ATCC). The human osteosarcoma cell line, HOS, was obtained from Dr. Richard Schwartz (Michigan State University, East Lansing, MI). Cell lines were maintained in Dulbecco's modified Eagle's medium (Invitrogen, Carlbad, CA) supplemented with 5% (v/v) fetal bovine serum, 5% newborn calf serum, and penicillin (50 units/mL) plus streptomycin (50 μg/mL) (Quality Biological, Gaithersburg, MD). The transfection vector, pNLNgoMIVITΔEnv.LUC, was prepared from pNLNgoMIVR-ΔEnv.HSA (Oh, J.; McWilliams, M. J.; Julias, J. G.; Hughes, S. H. Mutations in the U5 adjacent to the primer binding site affect tRNA cleavage by HIV-I RT in vivo. J. Virol, (in review), by replacing the HSA reporter gene (between Not I and Xhol) with a luciferase reporter gene between Notl and Xhol.

[0337] VSV-g -pseudotyped HIV was produced by transfection of 293 cells (Julias, J. G.; Boyer, P. L.; McWilliams, M. J.; Alvord, W. G.; and Hughes, S. H. Mutations at position 184 of human immunodeficiency virus type-1 reverse transcriptase affect virus titer and viral DNA synthesis. Virology 2004, 322, 13-21.) On the day prior to transfection, 293 cells were plated in 100 mm dishes at a density of 9x10⁵ cells per plate. 293 cells were transfected with 10 μg of pNLNgoMIVR^'ΔEnv.LUC and 3 μg of pHCMV-g (obtained from Dr. Jane Burns, University of California San Diego) using calcium phosphate precipitation. After 48 hours, virus-containing supernatants were harvested, clarified by low-speed centrifugation and filtration and diluted l-to-5 in preparation for infection assays. HOS cells were plated in 96 well luminescence cell culture plates at a density of 4000 cells in 100 μL per well the day prior to infection. On the day of infection, cells were pretreated with the target compounds for 3 h. Infections were carried out by adding 100 μL of virus-containing supernatants to each well and incubating for 48 h. Infectivity was measured via the luciferase reporter assay (Petropolous, C. J. et al. Antimicrob. Agents Chemother. 2000, 44, 920-928). Cells were lysed with 100 μL of Glo-Lysis Buffer (Promega, Madison, WI). Luciferase activity was measured by adding 100 μL of Steady-Glo reagent (Promega) directly to the lysed cells and measuring luminescence using a microplate reader. Activity was normalized to infections in the absence of target compounds. NFit (University of Texas, Galveston, Texas) was used to perform regression analysis on the data. EC₅₀ values were determined from the fit model. Inhibition of recombinant HIV-I integrase in vitro by hydrazide, amide, phthalimide and phthalhydrazide analog s in a metal-independent manner

[0338] Hydrazide, amide, phthalimide and phthalhydrazide analog s (Table 1-5) were tested for their ability to inhibit HIV-I integrase in vitro using 21 bp duplex that mimics viral LTR end. The IC₅₀ values obtained for the inhibition of HIV- 1 integrase in vitro by hydrazide, amide, phthalimide and phthalhydrazide analog s are summarized in Tables 1-5. Bis-salicylhydrazide (5a) exhibited potent but non-selective inhibition of both 3'-P and ST reactions in assays using purified recombinant IN with Mn²⁺ as cofactor. There was a striking loss of inhibitory potency was observed when the assays were performed using Mg²⁺ (entry 1, Table 1). A series of hydrazides was prepared in which a "right side" (2,3- dihydroxybenzoyl) group was combined with variety of "left side" components (8a - 8e, entries 2 - 6). Maintaining the left side salicyl group found in 5a gave inhibitor 8a (entry 2), whose inhibitory profile was unchanged from the parent 5a. In contrast, adding a 3-OH to the left side salicyl ring to provide the bis-(3,4-dihdroxybenzoyl)hydrazide (8b, entry 3) conferred good but non-selective 3'-P and ST inhibitory potencies with both Mg²⁺ as well as Mn²⁺ cofactors. Converting the left side 3-OH to a 3-OMe (8c, entry 4) had little effect on inhibitory potency in Mn²⁺, but reduced inhibitory potency in Mg²⁺ by slightly more than 20- fold. Extending the left side benzoyl ring system of 8a to a naphthoyl system (8d, entry 5) reduced inhibitory potency in Mn²⁺, and gave low but measurable inhibition in Mg²⁺. Removal of the hydroxyl in the left side aryl ring and introduction of a nitrogen to form a picalinoyl group (8e, entry 6) resulted in the lost of all inhibitory potency, except for very weak ST inhibition in Mn²⁺. This was very similar to the effects of completely removing the left side (13a, entry 10). Replacement of the left side hydrazide carbonyl in 8b with a hydrazone moiety (14a, entry 11) had little effect on inhibitory potency in Mn²⁺ but significantly reduced inhibitory potency in Mg²⁺. The corresponding 2,3-dimethoxybenzyl hydrazone (14b, entry 12) had reduced inhibitory potency. Introducing into the hydrazide N- N bond in 8b a 2-methylene spacer (9a, entry 7) or a 3-methylene spacer (9b, entry 8) reduced inhibitory potency in Mg²⁺. The 4-fluorobenzylamide analogue 9c (entry 9) showed ST inhibition only at the upper limits of the assay detection. Replacing the 2,3- dihydroxybenzoyl "right side" with a 2-hydroxy-3-methoxybenzoyl group and examining a similar series of left side- modified hydrazides (entries 13-18) generally resulted in little or no measurable inhibitory potency, except for 12a and 12b, which had good potency in assays using Mn²⁺ but not Mg²⁺ cofactor. Table 1. In Vitro Inhibition of Integrase 3'-P and ST Reactions in the Presence of Mn 2+

or Mg²⁺ Cofactor.

IC₅₀ (μM) (uM)

Entry No. Structure Metal Cofai Entry

=^ιor 3'-P ST No. Structure Metal Cofac :tor ₃,._p

ST

Mg²* >333 >333 Mg²* >333 >333

1 5a T l 1 x 10 13a X = U ^{H H} XJ Mn²* 36 1 8 ± 0 3 Mn²* >333 111 ± 27 ± 13

Mg²* 112 ± 28 68 ± 38 Mn²* >333 18 ± 5

Mg²* 245 ± 87 123 ± 16

Mn²* 26 ± 5 22 ± 7

Mg²⁺ >333 >333

7 9a X = Mg²* 85 ± 9 50 ± 13 16 13b X = Mn²⁺ >333 ?333

8 9b X = Mg²* 33 ± 4 31 ± 8 17 14c X = Mg²* >333 >333

9 9c X = ^■ _Fxr^Λ Mg²* >333 108 ± 12 18 15 X = _FJX^Λ Mg²* >333 ?333

Introduction of Carboxamide Conformational Constraint in the Hydrazide Series: iV- (l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-benzamides.

[0339] For inhibitors C-E to optimally undergo metal interactions as shown in the general structure i (Figure 1), the heteroatoms forming the metal chelating triads should be coplanar. The potency enhancement of ring nitrogens ortho to the carboxamide groups (shown in red, Figure 1) have been thought to be derived from facilitating such coplanarity (White, E. H.; Bursey, M. M. J. Org. Chem. 1966, 31, 1912-1917) For bicyclic inhibitors typified by the naphthyridine nucleus in C, a coplanar alignment of the carboxamide carbonyl was maintained by removing the ortho nitrogen and linking the carboxamide nitrogen to the parent bicycle through methylene or oxomethylene bridges to form conformationally- constrained tricyclic inhibitors (Jin, H. et al. Bioorg. Med. Chem. Lett. 2006, 16, 3989-3992; Verschueren, W. G.. J. Med. Chem. 2005, 48, 1930-1940.) A similar rationale was used for the bicyclic compounds of type G (Figure 1), which can be viewed as conformationally- constrained variants of the 2,3-dihdroxybenzoylhydrazides and amides presented in Table 1.

Table 2. In Vitro Inhibition of Integrase 3'-P and ST Reactions in the Presence of Mg²⁺ Cofactor.

kj

General Structure

Entry X = IC₅₀ (μM)

No. - Entry X = IC₅₀ (μM)

3'-P No.

ST 3'-P ST

22a >333 129 ± 38 22i 7.0 ± 1 3 1 2 ± 04

2 22b U t >333 293 ± 47 10 ²²i >333 >333

22c 42 11

22d xΛ_' > >3333 170 ± 105 12 221 78 ± 18 0 6 ± 02

22e V N-i 69 ± 23 21 ± 20 13 22m 6 7 ± 1 9 0 81 ± 0 35

22f J 186 ±47 53 ±18

7 22g 1 6 ± 0 8 021 ± 0 10 15 22o 87 ±29 54 ±12

22h 124 ± 32 41 ± 5 15 22p >333 >333

[0340] Examination of "left side" effects is the focus of compounds listed in Table 2, where poor inhibitory potencies resulted from unsubstituted (22a, entry 1) or 2-hydroxy mono- substituted (22b, entry 2) benzoylhydrazides. Addition of a second hydroxyl group provided a 50-fold enhancement in potency with greater than 50-fold selectivity for ST versus 3'-P (22c, entry 3). This contrasts with the absence of ST selectivity in the corresponding non- constrained analogue 8b (entry 3, Table 1). The 4-fluorobenzoylhydrazide (22d, entry 4) showed low inhibitor potency, with ST inhibition falling between the values shown by 22a and 22b. Constraining the 2,3-dihdroxybenzoyl left side in a fashion identical to the right side reduced ST inhibitory potency and decreased selectivity over 3'-P (22h, entry 8). Interestingly, the sulfonamide variant of this latter compound showed high inhibitory potency against both 3'-P and ST (22i, entry 9). The non-constrained bis-(3,4- dihydroxybenzenesulfonamide)-containing compound 22g (entry 7) showed good inhibitory potency against both 3'-P and ST reactions. Using a left side phthalimide motif to induce conformational constraint resulted in no measurable inhibition for the unsubstituted analogue 22j (entry 10) and moderate inhibitory potency for the 4-hydroxy derivative (22k). However, high inhibitory potency and greater than 100-fold ST selectivity resulted for the 4,5- dihydroxyphthalimide derivative 221 (entry 12). Equally high ST inhibitory potency but 10- fold less selectivity over 3'-P was observed for the isomeric 5,6-dihydroxyphthalimide (22m, entry 13).

Introduction of Carboxamide Conformational Constraint in the Benzamide Series.

[0341] The compounds 22 (Table 2) are all hydrazides intended to examine aspects of the previously reported inhibitor 5a. We prepared a parallel series of bicyclic analogues that would represent conformationally-constrained variants of the 2,3-dihydroxybenzamide nucleus (6, Figure 1). These bicycles are of two types, the 2,3-dihydro-6,7-dihydroxy-lH- isoindol-1-ones (24a - g) and the 4,5-dihydroxy-2-lH-isoindole-l,3(2H)-diones ("phthalimides" 27a - i), which differ in having either one or two oxo-groups in their isoindolyl hetero-rings, respectively (Table 3). As stated above, this approach has recently been applied to bicyclic nuclei such as the hydroxynaphthyridines, to yield tricyclic analogues (Jin, H. et al. Bioorg. Med. Chem. Lett. 2006, 16, 3989-3992; Verschueren, W. G. et al. J. Med. Chem. 2005, 48, 1930-1940)

2,3-Dihydro-6,7-dihydroxy- 1 H-isoindol- 1-ones.

[0342] In the 1 H-isoindol- 1 -one series an unsubstituted benzyl group provided good ST inhibitory potency and high selectivity over 3'-P (24a, entry 1, Table 3). Adding a methylene had little effect on ST inhibitory potency (24b, entry 2), however replacing the benzyl group with a 1-naphthyl system (24c, entry 3) increased 3'-P and ST inhibitory potencies by more than 5-fold and 10-fold, respectively. It has been reported in related series of compounds that adding a 4-fluoro substituent to the benzyl ring enhances inhibitory potency (for example, see Guare, J. P. et al. Bioorg. Med. Chem. Lett. 2006, 16, 2900-2904; Verschueren, W. G.; et al. J. Med. Chem. 2005, 48, 1930-1940). This has been attributed to the binding of the 4- flourophenyl group in a hydrophobic pocket present in the integrase^#DNA complex (Jin, H. et al. Bioorg. Med. Chem. Lett. 2006, 16, 3989-3992). However, in the present series, adding a 4-fluoro substituent had little effect (24d, entry 4). This contrasts sharply with the 3-chloro-4- fluoro substituted analogue (24e, entry 5), which exhibits significant enhancement of both 3'- P and ST inhibitory potencies relative to the 4-fluoro compound. This agrees with the previously reported beneficial effects of 3-chloro-4-fluoro substitution (Verschueren, W. G.; et al. J. Med. Chem. 2005, 48, 1930-1940).

[0343] During the early stages of DKA-based analogue development the 3- (phenylmethyl)phenyl group had shown utility in the design of high affinity inhibitors (Wai, J. S. et al. /. Med. Chem. 2000, 43, 4923-4926; Zhuang, L. et al. J. Med. Chem. 2003, 46, 453-456). However, in the present series this motif proved to be deleterious (24g, entry 7). Finally, the importance of the three oxygen atoms in the 2,3-dihydro-6,7-dihydroxy-lH- isoindol-1-ones is shown by the significant reduction in ST inhibitory potency of 24d incurred by either transposing the position of the oxo-group (28, entry 8) or by removing the 6-hydroxyl substituent (29, entry 9).

Table 3. In Vitro Inhibition of Integrase 3'-P and ST Reactions in the Presence of Mg²⁺ Cofactor.

IC₅₀ (μM)

Entry No. IC₅₀ (μM)

Structure Entry No. Structure

3'-P ST 3'-P ST

24a X = CT' >333 12 3 ± 5 6 10 27a X = -y 72 ± 23 039 ± 0 21

24b X = Gu -A- >333 28 ± 10 27b X = Ou, 157 ± 28 7 ± 3

24c X = 150 ± 36 0 72 ± 0 25

y

24d X = 282 ± 41 10 ± 4 13 27d X - = XT 8 ± 3 5 ± 2

24e X ^■ :xr- 13 ± 3 0 16 ± 008 14 27e X 14 ± 3 0 17 ± 006

24f X = _ Br Y^ 37 ± 12 13 ± 7 15 27f X = 2 8 ± 1 3 1 2 ± 06

24g X = 27 ± 6 0 64 ± 0 22

0 19 9

4,5-Dihydroxy-2-lH-isoindole-l,3(2H)-diones.

[0344] Adding a second oxo-group to the isoindole-1-one series (24) gave the isoindole- l,3(2H)-diones (27a - g). With a few exceptions (27c and e), this generally resulted in enhancement of 3'-P and ST inhibitory potencies (Table 3). Significant enhancements of ST inhibitory potencies resulted for 7V~substituents consisting of benzyl (27a, entry 10), 3- (bromomethyl)benzyl (27f, entry 15) and 3-(phenylmethyl)benzyl (27g, entry 16) groups. The high ST inhibitory potency of 27g was particularly noteworthy in light of the unexpected low potency of the corresponding isoindole-1-one analogue 24g. The lack of effect on the 3- chloro-4-fluorobenzyl-containing analogues 27e relative to 24e was also noteworthy, since this was in contrast to the large increase in 3'-P inhibitory potency for the corresponding A- fluorobenzyl-substituted analogue (27d) relative to 24d. Introducing a 7-amino group to the benzyl-substituted analogue 27a gave compound 31, which had good ST potency. Cellular Assays using HIV-I Based Vectors.

[0345] The antiviral effects of a select set of inhibitors was tested using HIV- 1 based vectors in cultured cells (Table 4). These studies showed that the bicyclic conformationally constrained analogues 24a, 24d and 27d exhibited sub-micromolar antiviral potencies. Elimination of the conformational constraint from 27d (compound 9c) or removal of the 4- fluoro-substituent (compound 27a) resulted in significant loss of antiviral potency. Interestingly, the 3-bromomethyl-substituted phthalimide 27f failed to exhibit good antiviral potency in spite of its low micromolar inhibition constant in extracellular IN assays. Hydrazides 5a, 8a and 8b all exhibited low micromolar antiviral effects. While the data for 8b was consistent with its potent inhibition of IN in extracellular assays employing Mg²⁺ cofactor, the antiviral effects observed for 5a and 5b potentially indicate cellular mechanisms of action independent of IN.

Table 4. Antiviral Potencies of Select Inhibitors in HIV-I Infected Cells.

Table 5. Structure-activity relationship of inhibition HIV IN and inhibition of HIV- induced cytopathic effect in cell culture for phthalimide derivatives.

All data represent mean values and standard deviations for at least three independent experiments.

Table 6. In vitro HIV-I inhibitory potencies of aryl and halobenzyl substituted compounds.

Comp. 3'-P ST

-Cl 24k 27k

72 ±34 3.5 ±1.1 13.7 ± 3.3 0.38 ± 0.1

O' Br 24I 264 ± 98 3.3 ± 1.2 27! 16.1 ±1.3 0.44 ±0.1

-I 24m 169 ±68 4.2 ±2.7 27m 85 ±35 1.1 ±0.4

Vc. 24uu 89 ± 15 0.10 ±0.02 27rr 18 ±4 0.12 ±0.02

27ss 111 ±13 9.8 ± 1.4

27vv 290 ±18 11.8 ±2.1

H l 24hh 27hh

>333 22.6 ±5.6 36.7 ±4.2 4.3 ±1.3

58.08 ±4.19 2.37 ±2.12

Table 7. In vitro HIV-I inhibitory potencies of monofluorobenzyl substituted compounds.

Com p. 3'-P ST

f\ 24i 27i

>333 24.59 ±5.26 68.08 ±3.45 2.39 ±1.48

11 ± 3 0.33 ± 0.19

58 ±20 0.93 ± 0.38

90 ±26 1.3 ±0.22

24t 17.9 ± 1.4 1.2 ±0.4 271 8.9* 1.2 0.31 ± 0.06

^F V J 24u >333 18.3 ±1.32 27u 54 ±9 3.5 ±1.4 Cl

-Cl 24r 27 r

36 ±17 1.7 ±1.1 25 ±16 1.7 ±0.8

<*^r V_Br ^24aa 33.5 ± 5.4 1.2 ±0.13 27 aa

11 ± 1.2 0.9 ±0.27

Table 8. Inhibitory potencies of difluorobenzyl substitutes inhibitors

24z 36.9 ±1.8 0.49 ±0.11 27z 16.3 ±3.5 0.27 ±0.08

^i=T^f= 24p 27p

333 26.58 ±4.33 60 ±7 0.23 ±0.18

27 ±1 0.14 ± 0.03

48 ±4 0.39 ±0.07

103 ±1 2.1 ±1.1

3.7 ± 0.89

>333 8.9 ± 2.5 27ff 34±2

78.8 ±31 0.38 ±0.17

Table 9. Inhibitory potencies of substitutes inhibitors

Comp. 3'-P ST Comp.

ST

F~\ y-F 24bb >333 25 ±6 27bb 158 ±16 1.76 ±0.4

27cc 18 ± 2 0.79 ±0.21

24dd >333 3.7 ±1.2 27dd _{54 i 4} 0.45 ±0.11

9.8± 1.5 058 ±0.17

24X 230 ±19 7.8 ± 2.7 ^27x 40.8 ± 6.6 1.25*0.46

300 ± 27 1.1 ±0.1

>333 3.17 ±0.64

Table 10. Antiviral Potencies of Select 2,3-dihydro-6,7-dihydroxy-lH-isoindol-l-one inhibitors in HIV-I Infected Cells

Table 11. Antiviral Potencies of Select 4,5-dihydroxy-lH-isoindole-l,3(2H)-dione inhibitors in HIV-I Infected Cells

[0346] Chelation of divalent Mg²⁺ is thought to be central to IN inhibition by several classes of agents, including diketoacids (A and B), 8-hydroxynaphthyridines (C) and 5- hydroxy-6-oxopyrimidinecarboxamides such as D. The bis-salicylhydrazide G is typical of an alternate family of IN inhibitors that have also been postulated to function by metal chelation. However, the hydrazide inhibitors exhibit high inhibitory potency only when Mn²⁺ is used as cofactor and they have little potency in the presence of Mg²⁺. The current study was undertaken to understand the mechanistic disparity between the bis-salicylhydrazides and other classes of metal chelators. For non-constrained hydrazides, potent inhibition in the presence of Mg²⁺ was found to require dihydroxybenzoyl substituents on both the right and left sides (for example, 8b). Elimination of a phenolic hydroxyl from one side either by removal (8a) or by conversion to a methyl ether (8c) or by removing a benzoyl carbonyl (14a) significantly reduced inhibitory potency in Mg²⁺, although good potency in Mn²⁺ could be retained. High selectivity of ST versus 3'-P found for many DKA inhibitors, was not observed in Mg²⁺ for the non-constrained hydrazides. However, good ST selectivity in Mg²⁺ could be achieved by conformational constraint of at least one benzoyl carbonyl using an isoindole ring structure (for example 22c and 221).

[0347] Conversion of the constrained dihydroxylated isoindole hydrazide series 22 to the corresponding amide series (24) generally resulted in good inhibitory potency in Mg²⁺ and ST selectivity without the need for metal-chelating functionality on the "left side" amide group. The arrangement of oxygen functionality on the "right side" dihydroxylated isoindole portion was critical, since all activity was lost on rearrangement of the carbonyl (28) or removal of a hydroxyl group (29). Carbonyl conformational constraint was important, as the constrained benzylamides 24a and 24d exhibited 10-fold higher potency than the non-constrained amide 9c. Adding a second carbonyl to yield the phthalimide series (27) in general enhanced inhibitory potency, including against 3'-P reactions. A striking example of potency enhancement is exemplified by the 3-(phenylmethyl)benzyl amide 27g, which exhibited greater than 250-fold higher ST inhibitory potency as compared to the isoindole congener 24g. The 4,5-dihydroxyphthalimide nucleus offers a structurally simple starting point for the further development of IN inhibitors.

[0348] All patents, patent applications, and other publications cited in this application, including published amino acid or polynucleotide sequences, are incorporated by reference in the entirety for all purposes. Any conflict between any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter. Likewise, any conflict between an art-understood definition of a word or phrase and a definition of the word or phrase as specifically taught in this specification shall be resolved in favor of the latter.

Claims

WHAT IS CLAIMED IS:

1. A compound having a formula selected from the group consisting of:

(III); and

wherein R* is H or -C|-C₈alkylaryl, R² is H, OH or -Ci-Cg alkoxy; each R^ is independently H or -Ci-C₈ alkyl; R⁴ is selected from the group consisting of: -Q-Cgalkylaryl, -C₃-C₂₄cycloalkyl, - alkylheteroaryl, -LN(R^4a)₂, or -LN=CH-aryl and -LN=CHheteraryl; each R^4a is independently selected from the group consisting of: H, -CO-aryl, - N=CH-aryl and -SO₂aryl; or is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(V); each R⁵ is independently selected from the group consisting of: haloC|-C₈alkyl-, -Ci- Qalkylaryl, -halo, -amino, -NHSO₂R⁵'¹, -N(SO₂R^5a)₂, OH, -C,-C₈alkoxy; heteroC,-C₈alkyl- -COR^5a and -NHCOR^5a; R^5a is Ci-Cgalkyl, aryl, 0R^5b or NHR^5b; R^5b is -C,-C₈alkyl or -NHN=CH-aryl; R⁶ is -CO-aryl ; -CH-aryl or -CH-C₃-C₂₄cycloalkyl ; Y¹ is CH₂, CO or SO₂; Y² is CH₂, CO or SO₂; Y³ is N, CH or CR⁵; L is a bond or a C2-C3alkenylene group; n is i, 2 or 3; and each of aryl, heterocyclyl, heteraryl or cycloalkyl is optionally substituted with from one to five substituents independently selected from the group consisting of: C_rC₈alkyl, haloQ- Cgalkyl-, -C, -Qalkylaryl, -halo, -amino, -NHSO₂R⁵³, -N(SO₂R^5a)₂, OH, -C,-C₈alkoxy; heteroC,- Cgalkyl-, heterocyclyl, -COC,-C₈alkyl, -CO₂C, -Cgalkyl; -COaryl and CO₂aryl; the dashed line indicates an optional bond; the wavy line indicates the point of attachment to the rest of the molecule; and pharmaceutically acceptable derivatives thereof.

2. The compound of claim 1, wherein the compound has formula II.

3. The compound of claim 1, wherein the compound has formula III.

4. The compound of claim 1, wherein the compound has formula IV.

5. The compound of claim 2, having the formula Ha:

6. The compound of claim 3, having the formula IHa:

7. The compound of claim 3, having the formula IHb:

8. The compound of any one of claims 1 to 7, wherein R⁶ is -CO-aryl.

9. The compound of any one of claims 1 to 7, wherein R⁶ is -CH-aryl.

10. The compound of any one of claims 1 to 7, wherein R⁶ is -CH-C₃-

C₂₄cycloalkyl.

11. The compound of any one of claims 1 to 10, wherein R⁴ is -C,-

C₈alkylaryl.

12. The compound of any one of claims 1 to 10, wherein R⁴ is -C₃-

C₂₄cycloalkyl.

13. The compound of any one of claims 1 to 10, wherein R is -LN(R )₂,

14. The compound of any one of claims 1 to 10, wherein R is -LN=CH- aryl.

15. The compound of any one of claims 1 to 10, wherein R ₄ i •s -

LN=CHheteraryl.

16. The compound of any one of claims 1 to 10, wherein R⁴ is -NHN=CH- Ph or -CH₂Ph.

17. The compound of claim 13, wherein R^^a has the formula VI:

wherein

Y² is CH₂, CO or SO₂; and each Y³ is N, CH or CR⁵.

18. The compound of any one of claims 13, wherein each R^4a is is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(Va) wherein each Y³ is N, CH or CR⁵.

19. The compound of any one of claims 13, wherein each R^4a is is taken together with the nitrogen to which each is attached to form a moiety of the formula:

(Vb) wherein each Y³ is N, CH or CR⁵.

20. The compound of any one of claims 13, wherein each R >⁴4^aa is is taken together with the nitrogen to which each is attached to form a moiety of the formula:

wherein Y³ is N, CH or CR⁵.

21. The compound of any one of claims 1 to 20, wherein R⁵ is independently selected from the group consisting of: -halo, -amino, -NHSO₂R⁵⁰, -N(SO₂R^5a)₂,, OH, -C_rC₈alkoxy, - COR^5a and -NHCOR^5a.

22. The compound of any one of claims 1 to 21, wherein Y¹ is CH₂.

23. The compound of any one of claims 1 to 21, wherein Y¹ is CO.

24. The compound of any one of claims 1 to 21, wherein Y¹ is SO₂.

25. The compound of any one of claims 1 to 21, wherein Y² is CH₂.

26. The compound of any one of claims 1 to 21, wherein Y² is CO.

27. The compound of any one of claims 1 to 21, wherein Y² is SO₂.

28. The compound of any one of claims 1 to 24, wherein L is a bond.

29. The compound of any one of claims 1 to 24, wherein L is a C2-

Cβalkenylene group.

30. The compound of any one of claims 1 to 29, wherein R is H.

31. The compound of any one of claims 1 to 29, wherein R¹ is -Q-

Qalkylaryl.

32. The compound of any one of claims 1 to 31, wherein R is H.

33. The compound of any one of claims 1 to 31, wherein R is OH.

34. The compound of any one of claims 1 to 31, wherein R is -Q-C₈ alkoxy.

35. The compound of any one of claims 1 to 34, wherein R³ is H.

36. The compound of any one of claims 1 to 34, wherein R³ is -C₁-C₈ alkyl.

37. The compound of any one of the preceding claims, wherein Q -C₈ alkyl is -Me or Et.

38. The compound of any one of the preceding claims, wherein aryl is phenyl.

39. The compound of any one of the preceding claims, wherein C₃- C₂₄cycloalkyl is cyclopropyl.

40. The compound of claim 1, selected from the group consisting of: 2,3-Dihydroxybenzoic Acid 2-(2-Hydroxybenzoyl)hydrazide; 2,3-Dihydroxybenzoic Acid 2-(2,3-Dihydroxybenzoyl)hydrazide; 2,3-Dihydroxybenzoic Acid 2-(2-Hydroxy-3-methoxybenzoyl)hydrazide; 2,3-Dihydroxybenzoic Acid 2-(3-Hydroxy-2-naphthoyl)hydrazide; 2,3-Dihydroxybenzoic Acid 2-Picalinoylhydrazide; N,N'-1 ,3-Ethanediylbis[2,3-dihydroxybenzamide]; N,N'-l,3-Propanediylbis[2,3-dihydroxybenzamide]; N-(4-Fluorobenzyl)-2,3-dihydroxybenzamide; 2-Hydroxy-3-methoxybenzoic Acid 2-(2-Hydroxy-3- methoxybenzoyl)hydrazide; 2-Hydroxy-3-methoxybenzoic Acid 2-(3-Hydroxy-2-naphthoyl)hydrazide; 2-Hydroxy-3-methoxybenzoic Acid 2-Picalinoylhydrazide; 2,3-Dihydroxybenzoic Acid Hydrazide; 2-Hydroxy-3-methoxybenzoic Acid Hydrazide; 2,3-Dihydroxybenzoic Acid [(2,3-Dihydroxyphenyl)methylene]hydrazide; 2,3-Dihydroxybenzoic Acid [(2,3-Dimethoxyphenyl)methylene]hydrazide; 2-Hydroxy-3-methoxybenzoic Acid [(4-Fluorophenyl)methylene]hydrazide; N-(4-Fluorobenzyl)-2-hydroxy-3-methoxybenzamide; 1 ,2-Dimethoxy-3-(methoxymethyl)benzene; 2,3-Dimethoxy-6-(methoxymethyl)benzoic Acid Methyl Ester; 6-(Chloromethyl)-2,3-dimethoxybenzoic Acid Methyl Ester; 2-Amino-2,3-dihydro-7,8-dimethoxy-lH-isoindol-l-one; N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-benzamide; N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-(2- hydroxy)benzamide; N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-(2,3- dimethoxy)benzamide; N-(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-N-(4-fluorobenzoyl)- 4-flourobenzamide; N-(1 ,3-Dihydro-6,7-dimethoxy- 1 -oxo-2H-isoindol-2-yl)-N-(4- fluorobenzenesulfonyl)-4-flourobenzesulfonamide; N-(1 ,3-Dihydro-6,7-dimethoxy- 1 -oxo-2H-isoindol-2-yl)-N-(3 ,4- dimethoxybenzenesulfonyl)-3,4-dimethoxybenzesulfonamide; ό.ό'JJ'-Tetramethoxy-P^'-bi^H-isoindolel-iαXSH^'^-dione; N-[(l,3-Dihydro-6,7-dimethoxy-l-oxo-2H-isoindol-2-yl)-N'-[(6,7-dimethoxy- 1,1 -dioxido- 1 ,2-benzisothiazol-2(3H)-yl)] Hydrazide; 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-isoindole- l,3(2H)-dione; 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4- hydroxyisoindole-l,3(2H)-dione; 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5- dimethoxyisoindole-l,3(2H)-dione; 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5- dimethoxyisoindole-l,3(2H)-dione; 2-[l,3-Dihydro-6,7-dimethoxy-3-oxo-2H-isoindol-2-yl]-lH-4-fluoroisoindole- l,3(2H)-dione; 2,3-Dihydro-6,7-dimethoxy-2-[(2,3-dihydroxyphenylmethylene)amino]-lH- Isoindol-1-one; 2,3-Dihydro-6,7-dimethoxy-2-[(4-fluorophenylmethylene)amino]-lH- Isoindol-1-one; N-( 1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-benzamide; N-(1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-(2- hydroxy)benzamide; N-(1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-(2,3- dihydroxy)benzamide; N-( 1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-4-flourobenzamide; N-(1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-N-(4- fluorobenzenesulfonyl)-4-flourobenzesulfonamide (22e) and N-(l,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-4-flourobenzesulfonamide; N-(1 ,3-Dihydro-6,7-dihydroxy- 1 -oxo-2H-isoindol-2-yl)-N-(3 ,4- dihydroxybenzenesulfonyl)-3,4-dihydroxybenzesulfonamide; 6,6',7,7'-Tetrahydroxy-[2,2^l-bi-2H-isoindole]-l,l'(3H,3Η)-dione; N-[(l,3-Dihydro-6,7-dihydroxy-l-oxo-2H-isoindol-2-yl)-N'-[(6,7-dihydroxy- l,l-dioxido-l,2-benzisothiazol-2(3H)-yl)] Hydrazide; 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-isoindole-l,3(2H)- dione; 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-3- hydroxyisoindole- 1 ,3(2H)-dione; 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-3,4- dihydroxyisoindole- 1 ,3(2H)-dione; 2-[l,3-Dihydro-6,7-dihydoxy-3-oxo-2H-isoindol-2-yl]-lH-4,5- dihydroxyisoindole- 1 ,3(2H)-dione; 2-[l,3-Dihydro-6,7-dihydroxy-3-oxo-2H-isoindol-2-yl]-lH-4-fluoroisoindole- l,3(2H)-dione; 2,3-Dihydro-6,7-dihydroxy-2-[(2,3-dihydroxyphenylmethylene)amino]-lH- Isoindol-1-one; 2,3-Dihydro-6,7-dihydroxy-2-[(4-fluorophenylmethylene)amino]-lH-Isoindol- 1-one; 2,3-Dihydro-6,7-dimethoxy-2-phenylmethyl-H-isoindol- 1-one; 2,3-Dihydro-6,7-dimethoxy-2-(2-phenylethyl)-H-isoindol- 1 -one; 2,3-Dihydro-6,7-dimethoxy-2-( 1 -naphthylmethyl)-H-isoindol- 1 -one; 2,3-Dihydro-6,7-dimethoxy-2-(4-fluorophenylmethyl)-H-isoindol- 1 -one; 2,3-Dihydro-6,7-dimethoxy-2-[(3-chloro-4-fluorophenyl)methyl]-H-isoindol- 1-one; 2,3-Dihydro-6,7-dimethoxy-2-[3-((hydroxymethyl)phenyl)methyl]-H- isoindol-1-one; 2,3-Dihydro-6,7-dimethoxy-2-[3-((phenylmethyl)phenyl)methyl]-H-isoindol- 1-one; 2,3-Dihydro-6,7-dihydroxy-2-ρhenylmethyl-H-isoindol- 1 -one; 2,3-Dihydro-6,7-dihydroxy-2-(2-phenylethyl)-H-isoindol-l-one; 2,3-Dihydro-6,7-dihydroxy-2-(l-naphthylmethyl)-H-isoindol- 1-one; 2,3-Dihydro-6,7-dihydroxy-2-(4-fluorophenylmethyl)-H-isoindol- 1 -one; 2,3-Dihydro-6,7-dihydroxy-2-[(3-chloro-4-fluorophenyl)methyl]-H-isoindol- 1-one; 2,3-Dihydro-6,7-dihydroxy-2-[3-((bromomethyl)phenyl)methyl]-H-isoindol- 1 - one; 2,3-Dihydro-6,7-dihydroxy-2-[3-((phenylmethyl)phenyl)methyl]-H-isoindol- 1-one; 4,5-Dihydroxy-2-[(3-fluorophenyl)methyl]-H-isoindol- 1-one; 4,5-DihydiOxy-2-[(2-fluorophenyl)methyl]-H-isoindol- 1-one; 102 4,5-Dihydroxy-2-[(3-(azidomethyl)phenyl)methyl]-H-isoindol-l-one;

103 4,5-Dihydroxy-2-[(3-chlorophenyl)methyl]-H-isoindol- 1-one;

104 4,5-Dihydroxy-2-[(3-bromophenyl)methyl]-H-isoindol-l-one;

105 4,5-Dihydroxy-2-[(3-iodophenyl)methyl]-H-isoindol-l-one;

106 4,5-Dihydroxy-2-[(3,4-difluorophenyl)methyl]-H-isoindol-l-one;

107 4,5-Dihydroxy-2-[(3,5-difluorophenyl)methyl]-H-isoindol-l-on;

108 4,5-Dihydroxy-2-[(2,5-difluorophenyl)methyl]-H-isoindol-l-one;

109 4,5-Dihydroxy-2- [(2,6-difluorophenyl)methyl] -H-isoindol- 1 -one;

110 4,5-Dihydroxy-2-[(2-fluoro-3-chlorophenyl)methyl]-H-isoindol- 1 -one;

111 2-(2-chloro-4-fluorobenzyl)-4,5-dihydroxyisoindol- 1 -one;

112 2-(5-chloro-2-fluorobenzyl)-4,5-dihydroxyisoindol-l-one;

113 2-(2-chloro-6-fluorobenzyl)-4,5-dihydroxyisoindol- 1 -one;

114 2-(3-fluoro-4-methylbenzyl)-4,5-dihydroxyisoindol-l-one;

115 2-(4-fluoro-3-methylbenzyl)-4,5-dihydroxyisoindol- 1 -one;

116 4,5-dihydroxy-2-(perfluorobenzyl)isoindol- 1 -one;

117 2-(2,3-difluorobenzyl)-4,5-dihydroxyisoindol-l-one;

118 2-(2,4-difluorobenzyl)-4,5-dihydroxyisoindol- 1-one;

119 2-(2,4-difluorobenzyl)-4,5-dihydroxyisoindol- 1 -one;

120 4,5-dihydroxy-2-(2,3,6-trifluorobenzyl)isoindol-l-one;

121 4,5-dihydroxy-2-(2,3,4-trifluorobenzyl)isoindol- 1-one;

122 4,5-dihydroxy-2-(2,4,5-trifluorobenzyl)isoindol- 1-one;

123 4,5-dihydroxy-2-(3,4,5-trifluorobenzyl)isoindol- 1-one;

124 2-(2-chloro-3,6-difluorobenzyl)-4,5-dihydroxyisoindol-l-one;

125 2-(2-chloro-3,6-difluorobenzyl)-4,5-dihydroxyisoindol- 1-one; 126 2-(2,3-dihydro-lH-inden-2-yl)-4,5-dihydroxyisoindol-l-one;

127 2-(3-benzoylbenzyl)-4,5-dihydroxyisoindol-l-one;

128 4,5-dihydroxy-2-(3-(3-(trifluoromethyl)-3H-diazirin-3-yl)benzyl)isoindol-l-

129 one;

130 4,5-dihydroxy-2-(pyridin-2-ylmethyl)isoindol-l-one;

131 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindol-l-one;

132 (S)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindol-l-

133 one;

134 (R)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindol-l-

135 one;

136 (S)-2-((2,3-dihydro- 1 H-inden- 1 -yl)methyl)-4,5-dihydroxyisoindol- 1 -one;

137 (R)-2-((2,3-dihydro-l H-inden- l-yl)methyl)-4,5-dihydroxyisoindol-l -one;

138 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindol-l-one;

139 4,5-Dihydroxy-2-[(2-aminophenyl)methyl]-H-isoindol-l-one;

140 N-(2-((4,5-dihydroxy-l-oxoisoindolin-2-yl)methyl)phenyl)-N-

141 (methylsulfonyl)methanesulfonamide;

142 N-(2-((4,5-dihydroxy-l-oxoisoindolin-2-yl)methyl)

143 phenyl)methanesulfonamide;

144 4,5-Dimethoxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione;

145 4,5-Dimethoxy-2-[(2-phenyl)ethyl]-lH-isoindole-l,3(2H)-dione;

146 4,5-Dimethoxy-2-[(l-naphthyl)methyl]-lH-isoindole-l,3(2H)-dione;

147 4,5-Dimethoxy-2-[(4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

148 4,5-Dimethoxy-2-[(3-chloro-4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-

149 dione;

150 4,5-Dimethoxy-2-[(3-(phenylmethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-

151 dione;

152 4,5-Dimethoxy-2-[(3-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione; 153 4,5-Dimethoxy-2-[(3-(hydroxymethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-

154 dione;

155 4,5-Hydroxy-2-(phenylmethyl)-lH-isoindole-l,3(2H)-dione;

156 4,5-Dihydroxy-2-[(2-phenyl)ethyl]- lH-isoindole- 1 ,3(2H)-dione;

157 4,5-Dihydroxy-2-[(l-naphthyl)methyl]-lH-isoindole-l,3(2H)-dione;

158 4,5-Dihydroxy-2-[(4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

159 4,5-Dihydroxy-2-[(3-chloro-4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-

160 dione;

161 4,5-Dihydroxy-2-[(3-(bromomethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-

162 dione;

163 4,5-Dihydroxy-2-[(3-(hydroxymethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-

164 dione;

165 4,5-Dihydroxy-2-[(3-(phenylmethyl)phenyl)methyl]-lH-Isoindole-l,3(2H)-

166 dione;

167 4,5-Dihydroxy-2-[(3-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

168 4,5-Dihydroxy-2-[(2-fluorophenyl)methyl]- lH-isoindole- 1 ,3(2H)-dione;

169 4,5-Dihydroxy-2-[(3-(azidomethyl)phenyl)methyl]-lH-isoindole-l,3(2H)-

170 dione;

171 4,5-Dihydroxy-2-[(3-chlorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

172 4,5-Dihydroxy-2-[(3-bromophenyl)methyl]-lH-isoindole~l,3(2H)-dione;

173 4,5-Dihydroxy-2-[(3-iodophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

174 4,5-Dihydroxy-2-[(3, 4-difluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

175 4,5-Dihydroxy-2- [(3 ,5-difluorophenyl)methyl] - 1 H-isoindole- 1 ,3 (2H)-dione;

176 4,5-Dihydroxy-2-[(2,5-difluorophenyl)methyl]- lH-isoindole- 1 ,3(2H)-dione;

177 4,5-Dihydroxy-2-[(2,6-difluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

178 4,5-Dihydroxy-2-[(2-fluoro-3-chlorophenyl)methyl]-lH-isoindole-l,3(2H)-

179 dione;

180 2-(2-chloro-4-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione;

181 2-(5-chloro-2-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione; 182 2-(2-chloro-6-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione; 183 2-(3-fluoro-4-methylbenzyl)-4,5-dihydroxyisoindoline-l,3-dione; 184 2-(4-fluoro-3-methylbenzyl)-4,5-dihydroxyisoindoline- 1,3-dione; 185 4,5-dihydroxy-2-(perfluorobenzyl)isoindoline- 1 ,3-dione; 186 2-(2,3-difluorobenzyl)-4,5-dihydroxyisoindoline- 1 ,3-dione; 187 2-(2,4-difluorobenzyl)-4,5-dihydroxyisoindoline- 1,3-dione; 188 2-(2,4-difluorobenzyl)-4,5-dihydroxyisoindoline- 1,3-dione; 189 4,5-dihydroxy-2-(2, 3, 6-trifluorobenzyl)isoindoline- 1,3-dione; 190 4,5-dihydroxy-2-(2,3,4-trifluorobenzyl)isoindoline-l,3-dione; 191 4,5-dihydroxy-2-(2,4,5-trifluorobenzyl)isoindolin-l,3-dione; 192 4,5-dihydroxy-2-(3, 4, 5-trifluorobenzyl)isoindoline- 1,3-dione; 193 2-(2-chloro-3,6-difluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione; 194 2-(2-chloro-3,6-difluorobenzyl)-4,5-dihydroxyisoindoline- 1,3-dione; 195 2-(2,3-dihydro-lH-inden-2-yl)-4,5-dihydroxyisoindoline- 1,3-dione; 196 2-(3-benzoylbenzyl)-4,5-dihydroxyisoindoline- 1 ,3-dione;

197 4,5-dihydroxy-2-(3-(3-(trifluoromethyl)-3H-diazirin-3-yl)benzyl)isoindoline- 198 1,3-dione;

199 4,5-dihydroxy-2-(pyridin-2-ylmethyl)isoindoline- 1 ,3-dione; 200 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindoline- 1 ,3-dione;

201 (S)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindoline- 202 1,3-dione;

203 (R)-4,5-dihydroxy-2-((l,2,3,4-tetrahydronaphthalen-2-yl)methyl)isoindoline- 204 1 ,3-dione;

205 (S)-2-((2,3-dihydro- lH-inden- 1 -yl)methyl)-4,5-dihydroxyisoindoline- 1 ,3- 206 dione; 207 (R)-2-((2,3-dihydro-lH-inden-l-yl)methyl)-4,5-dihydroxyisoindoline-l,3-

208 dione;

209 4,5-dihydroxy-2-(naphthalen-2-ylmethyl)isoindoline- 1 ,3-dione;

210 2,3-Dihydro-4,5-dihydroxy-2-(4-fluorophenylmethyl)-H-isoindol-l-one;

211 4-Hydroxy-2-[(4-fluorophenyl)methyl]-lH-isoindole-l,3(2H)-dione;

212 4- Amino-6,7-dimethoxy-2-(phenylmethyl)- 1 H-isoindole- 1 ,3(2H)-dione; and

213 4-Amino-6,7-dihydroxy-2-(phenylmethyl)- 1 H-isoindole- 1 ,3 (2H)-dione.

1 41. The compound of claim 1, selected from the group consisting of:

2 2-(3-chlorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

3 2-(3-bromobenzyl)-6,7-dimethoxyisoindolin-l-one;

4 2-(3-iodobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

5 2-(3,4-dichlorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

6 6,7-dimethoxy-2-(naphthalen-2-ylmethyl)isoindolin- 1 -one;

7 (S)-6,7-dimethoxy-2-( 1 ,2,3,4-tetrahydronaphthalen- 1 -yl)isoindolin- 1 -one;

8 (R)-6,7-dimethoxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindolin-l-one;

9 (S )-2-(2,3 -dihydro- 1 H-inden- 1 -yl)-6,7 -dimethoxyisoindolin- 1 -one;

10 (R)-2-(2,3'dihydro-lH-inden-l-yl)-6,7-dimethoxyisoindolin-l-one;

11 2-(2,3-dihydro-lH-inden-2-yl)-6,7-dimethoxyisoindolin-l-one;

12 2-(2-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one;

13 2-(3-fluorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

14 2-(3-chloro-2-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one;

15 2-(4-chloro-3 -fluorobenzyl)-6 ,7-dimethoxyisoindolin- 1 -one;

16 2-(2-chloro-4-fluorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

17 2-(5-chloro-2-fluorobenzyl)-6,7-dimethoxyisoindolin-l-one;

18 2-(2-chloro-6-fluorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

19 2-(4-fluoro-3-methylbenzyl)-6,7-dimethoxyisoindolin-l-one;

20 2-(3-fluorσ-4-methylbenzyl)-6,7-dimethoxyisoindolin- 1 -one;

21 2-(3-bromo-4-fluorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

22 2-(2,3-difluorobenzyl)-6,7-dimethoxyisoindolin- 1 -one;

23 2-(2,4-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one;

24 2-(2,5-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one;

25 2-(3 ,4-difluorobenzyl)-6 ,7-dimethoxyisoindolin- 1 -one; 2-(3,5-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one; 2-(2,6-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one; 2-(2-chloro-3,6-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one; 2-(5-chloro-2,4-difluorobenzyl)-6,7-dimethoxyisoindolin-l-one; 6,7-dimethoxy-2-(2,3,6-trifluorobenzyl)isoindolin- 1 -one; 6,7-dimethoxy-2-(2,3,4-trifluorobenzyl)isoindolin-l-one; 6,7-dimethoxy-2-(2,4,5-trifluorobenzyl)isoindolin-l-one; 6,7-dimethoxy-2-(3,4,5-trifluorobenzyl)isoindolin- 1 -one; 6,7-dimethoxy-2-(perfluorobenzyl)isoindolin- 1 -one; 2-(3,4-dichlorobenzyl)-6,7-dihydroxyisoindolin-l-one; (S)-6,7-dihydroxy-2-( 1 ,2,3,4-tetrahydronaphthalen- 1 -yl)isoindolin- 1 -one; (R)-6,7-dihydroxy-2-( 1 ,2,3 ,4-tetrahydronaphthalen- 1 -yl)isoindolin- 1 -one; (S)-2-(2,3-dihydro-lH-inden-l-yl)-6,7-dihydroxyisoindolin-l-one; (R)-2-(2,3-dihydro-lH-inden-l-yl)-6,7-dihydroxyisoindolin-l-one; 2-(4-chloro-3-fluorobenzyl)-6,7-dihydroxyisoindolin-l-one; 2-(5-chloro-2,4-difluorobenzyl)-6,7-dihydroxyisoindolin-l-one; (S)-6,7-dihydroxy-2-( 1 -phenyl ethyl)isoindolin- 1 -one; (R)-6,7-dihydroxy-2-( 1 -phenylethyl)isoindolin- 1 -one; (S)-2-(l-(4-fluorophenyl)ethyl)-6,7-dihydroxyisoindolin-l-one; (R)-2-(l-(4-fluorophenyl)ethyl)-6,7-dihydroxyisoindolin-l-one; (S)-6,7-dihydroxy-2-( 1 -(naphthalen- 1 -yl)ethyl)isoindolin- 1 -one; (R)-6,7-dihydroxy-2-(l-(naphthalen-l-yl)ethyl)isoindolin-l-one; (S)-6,7-dihydroxy-2-(l-(naphthalen-2-yl)ethyl)isoindolin-l-one; (R)-6,7-dihydroxy-2-(l-(naphthalen-2-yl)ethyl)isoindolin-l-one; 2-(3-benzoylbenzyl)-4-bromo-6,7-dihydroxyisoindolin- 1 -one; 2-(4-benzoylbenzyl)-6,7-dihydroxyisoindolin- 1-one; 6,7-dihydroxy-2-(3-(3-(trifluoromethyl)-3H-diazirin-3-yl)benzyl)isoindolin-l- one; 2-(3-chlorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione; 2-(3-bromobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione; 2-(3-iodobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione; 2-(3,4-dichlorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione; 4,5-dimethoxy-2-(naphthalen-2-ylmethyl)isoindoline- 1 ,3-dione; (S)-4,5-dimethoxy-2-( 1 ,2,3,4-tetrahydronaphthalen- 1 -yl)isoindoline- 1,3- dione; (R)-4,5-dimethoxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindoline-l,3- dione;

(S)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dimethoxyisoindoline-l,3-dione;

(R)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2,3-dihydro-lH-inden-2-yl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2-fluorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione (26bbb)

2-(4-chloro-3-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2-chloro-4-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(5-chloro-2-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2-chloro-6-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(3-chloro-2-fluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(3-fluoro-4-methylbenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(4-fluoro-3-methylbenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(3-bromo-4-fluorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione;

2-(2,3-difluorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione;

2-(2,4-difluorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione ;

2-(2,5-difluorobenzyl)-4,5-dimethoxyisoindoline- 1 ,3-dione;

2-(3,4-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(3,5-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2,6-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(2-chloro-3,6-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

2-(5-chloro-2,4-difluorobenzyl)-4,5-dimethoxyisoindoline-l,3-dione;

4,5-dimethoxy-2-(2,3,6-trifluorobenzyl)isoindoline-l,3-dione;

4,5-dimethoxy-2-(2,3,4-trifluorobenzyl)isoindoline-l,3-dione;

4,5-dimethoxy-2-(2,4,5-trifluorobenzyl)isoindoline-l,3-dione;

4,5-dimethoxy-2-(3,4,5-trifluorobenzyl)isoindoline-l,3-dione;

4,5-dimethoxy-2-(perfluorobenzyl)isoindoline- 1 ,3-dione;

2-(3,4-dichlorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione;

(S)-4,5-dihydroxy-2-(l,2,3,4-tetrahydronaphthalen-l-yl)isoindoline-l,3-dione;

(R)-4,5-dihydroxy-2-(l ,2,3,4-tetrahydronaphthalen-l-yl)isoindoline- 1 ,3-dione;

(S)-2-(2,3-dihydro- lH-inden- 1 -yl)-4,5-dihydroxyisoindoline- 1 ,3-dione; 92 (R)-2-(2,3-dihydro-lH-inden-l-yl)-4,5-dihydroxyisoindoline-l,3-dione;

93 2-(4-chloro-3-fluorobenzyl)-4,5-dihydroxyisoindoline-l,3-dione;

94 2-(5-chloro-2,4-difluorobenzyl)-4,5-dihydroxyisoindoline- 1 ,3-dione; and

95 2-(4-benzoylbenzyl)-4,5-dihydroxyisoindoline- 1 ,3-dione.

1 42. A method for inhibiting retrovirus proliferation, comprising the step of

2 contacting a cell, which is infected with a retrovirus or at risk of being infected with a

3 retrovirus, with an effective amount of a compound of formula I or II:

(I);

6 wherein R^ is H or -CrQalkylaryl,

7 R² is H, OH or -C₁-C₈ alkoxy;

8 each R-^> is independently H or -Q-Cg alkyl;

9 R⁴ is selected from the group consisting of: -C_rC₈alkylaryl, -C₃-C₂₄cycloalkyl, -

10 alkylheteroaryl, -LN(R^4a)₂, or -LN=CH-aryl and -LN=CHheteraryl;

11 each R^4a is independently selected from the group consisting of: H, -CO-aryl, -

12 N=CH-aryl and -SC^aryl; or is taken together with the nitrogen to which each is attached to form a

13 moiety of the formula:

14 (V); each R⁵ is independently selected from the group consisting of: haloC_rC₈alkyl-, -Q-

Qalkylaryl, -halo, -amino, -NHSO₂R , -N(SO₂R )₂,, OH, -C,-C₈alkoxy; heteroC,-C₈alkyl- -COR >5a and -NHCOR^M;

R^5a is aryl, 0R^5b or NHR^5b;

R^5b is -C-Qalkyl or -NHN=CH-aryl;

R⁶ is -CO-aryl; -CH-aryl or -CH-C₃-C₂₄cycloalkyl;

Y¹ is CH₂, CO or SO₂;

Y² is CH₂, CO or SO₂;

Y³ is N, CH or CR⁵;

L is a bond or a C2-C3alkenylene group; n is 1, 2 or 3; and each of aryl, heterocyclyl, heteraryl or cycloalkyl is optionally substituted with from one to five substituents independently selected from the group consisting of: Ci-Csalkyl, haloCi- C₈alkyl-, -C,-C₈alkylaryl, -halo, -amino, -NHSO₂R^5a, -N(SO₂R^5a)₂, OH, -C,-C₈alkoxy; heteroC,- Cgalkyl-, heterocyclyl, -COC,-C₈alkyl, -CO₂C,-C₈arkyl; -COaryl and CO₂aryl; the dashed line indicates an optional bond; the wavy line indicates the point of attachment to the rest of the molecule; and pharmaceutically acceptable derivatives thereof.

43. A method for inhibiting retrovirus proliferation, comprising the step of contacting a cell, which is infected with a retrovirus or at risk of being infected with a retrovirus, with an effective amount of a compound of any one of claims 1 to 40.

44. The method of claim 42 or 43, wherein the contacting step is performed in vitro.

45. The method of claim 42 or 43, wherein the cell is a part of a living animal.

46. The method of claim 45, wherein the animal is a human.

47. The method of claim 42 or 43, wherein the retrovirus is HIV-I .